microRNA

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Diagram of miRNA action with mRNA

The stem-loop secondary structure of a pre-microRNA from Brassica oleracea.

A microRNA (abbr. miRNA) is a small non-coding RNA molecule (ca. 22 nucleotides) found in plants, animals, and some viruses, which functions in transcriptional and post-transcriptional regulation of gene expression.[1] Encoded byeukaryotic nuclear DNA in plants and animals and by viral DNA in certain viruses whose genome is based on DNA, miRNAs function via base-pairing with complementary sequences within mRNA molecules, usually resulting in gene silencing via translational repression or target degradation.[2][3] The human genome may encode over 1000 miRNAs,[4][5]which may target about 60% of mammalian genes[6][7] and are abundant in many human cell types.[8] miRNAs are well conserved in eukaryotic organisms and are thought to be a vital and evolutionarily ancient component of genetic regulation.[9][10][11][12] While core components of the microRNA pathway are conserved between plants andanimals, miRNA repertoires in the two kingdoms appear to have evolved independently with different modes of function.[13] Plant miRNAs usually have perfect or near-perfect pairing with their messenger RNA targets and induce gene repression through degradation of their target transcripts.[14][15] Plant miRNAs may bind their targets in both coding regions and untranslated regions.[15] In contrast, animal miRNAs are able

to recognize their target mRNAs by as little as 6-8 nucleotides (the seed region) at the 5' end of an animal miRNA.[6][16] Combinatorial regulation is a feature of miRNA regulation. A given miRNA may have multiple different mRNA targets, and a given target might similarly be targeted by multiple miRNAs.[17][18] The first miRNAs were characterized in the early 1990s.[19] However, miRNAs were not recognized as a distinct class of biological regulators with conserved functions until the early 2000s. Since then, miRNA research has revealed multiple roles in negative regulation (transcript degradation and sequestering, translational suppression) and possible involvement in positive regulation (transcriptional and translational activation). By affecting gene regulation, miRNAs are likely to be involved in most biological processes.[20][21][22][23][24][25][26] Different sets of expressed miRNAs are found in different cell types and tissues.[27] Aberrant expression of miRNAs has been implicated in numerous disease states, and miRNA-based therapies are under investigation.[28][29][30][31] Estimates of the average number of unique messenger RNAs that are targets for repression by a typical microRNA vary, depending on the method used to make the estimate.[32] While a 2004 estimate was that the mean number of target mRNAs for a typical microRNA is only 7,[33] later estimates were higher. Krek et al.[34] found that vertebrate microRNAs target, on average, roughly 200 transcripts each. Selbach et al. [35] and Baek et al.[36] indicated that a single miRNA may repress the production of hundreds of proteins, but that this repression often is relatively mild (less than 2-fold).

History[edit]
MicroRNAs were discovered in 1993 by Victor Ambros, Rosalind Lee and Rhonda Feinbaum during a [19] study of the gene lin-14 in C. elegans development. They found that LIN-14 protein abundance was regulated by a short RNA product encoded by the lin-4 gene. A 61-nucleotide precursor from the lin4 gene matured to a 22-nucleotide RNA that contained sequences partially complementary to multiple sequences in the 3' UTR of the lin-14 mRNA. This complementarity was both necessary and sufficient to inhibit the translation of the lin-14 mRNA into the LIN-14 protein. Retrospectively, the lin-4 small RNA was the first microRNA to be identified, though at the time, it was thought to be a nematode idiosyncrasy. Only in 2000 was a second RNA characterized: let-7, which repressed lin-41, lin-14, lin-28, lin-42, and daf12 expression during developmental stage transitions in C. elegans. let-7 was soon found to be [37][38] conserved in many species, indicating the existence of a wider phenomenon.

Nomenclature[edit]
Under a standard nomenclature system, names are assigned to experimentally confirmed miRNAs before [39][40] publication of their discovery. The prefix "mir" is followed by a dash and a number, the latter often indicating order of naming. For example, mir-123 was named and likely discovered prior to mir-456. The uncapitalized "mir-" refers to the pre-miRNA, while a capitalized "miR-" refers to the mature form. miRNAs with nearly identical sequences except for one or two nucleotides are annotated with an additional lower case letter. For example, miR-123a would be closely related to miR-123b. Pre-miRNAs that lead to 100% identical mature miRNAs but that are located at different places in the genome are indicated with an additional dash-number suffix. For example, the pre-miRNAs hsa-mir-194-1 and hsa-mir-194-2 lead to an

they are denoted with a -3p or -5p suffix. this provides a means for coupled regulation [45] of miRNA and protein-coding gene. The majority of the characterized miRNA genes are intergenic or oriented antisense to neighboring genes [41][41][42][43][44] and are therefore suspected to be transcribed as independent units. miR-123 and miR-123* would share a pre-miRNA hairpin. Biogenesis[edit] MicroRNAs are produced from either their own genes or from introns. found in a sense orientation.g. However. and thus usually are regulated together . When relative expression levels are known. When two mature microRNAs originate from opposite arms of the same pre-miRNA. e. Other common prefixes include 'v' for viral (miRNA encoded by a viral genome) and 'd' for Drosophila miRNA (a fruit fly commonly studied in genetic research). in some cases a microRNA gene is transcribed together with its host gene. this distinction was also made with 's' (sense) and 'as' (antisense)). though not exclusively.identical mature miRNA (hsa-miR-194) but are located in different regions of the genome. As much as 40% of miRNA genes may lie in the introns of protein [46] and non-protein coding genes or even inexons of long nonprotein-coding transcripts. but more miR-123 would be found in the cell.. an asterisk following the name indicates an miRNA expressed at low levels relative to the miRNA in the opposite arm of a hairpin. For example. Species of origin is designated with a three-letter prefix. These are [47][48] usually. hsa-miR-123 is a human (Homo sapiens) miRNA and oar-miR-123 is a sheep (Ovis aries) miRNA. A video of this process can be found here. (In the past.

a protein that cuts [54] RNA. enzymes known as adenosine deaminases acting on RNA (ADARs) catalyzeadenosine to inosine (A to I) transitions. especially those with upstream Alu sequences. When a stem-loop precursor is found [47] in the 3' UTR. [46][49][50] Transcription[edit] miRNA genes are usually transcribed by RNA polymerase II (Pol II). It is often termed as a pre-miRNA (precursor-miRNA). DGCR8 orients the catalytic RNase III domain of Drosha to liberate hairpins from pri-miRNAs by cleaving RNA about eleven nucleotides from the hairpin base (two helical RNA turns into the stem). [42][52] Nuclear processing[edit] A single pri-miRNA may contain from one to six miRNA precursors. Most commonly. polyadenylated with [42][47] multiple adenosines (a poly(A) tail). to form the "Microprocessor" complex. The DNA template is not the final word on mature miRNA production: 6% of human miRNAs show RNA editing (IsomiRs). by changing the seed region of miR[56] 376 in the central nervous system). leading to degradation by the ribonuclease Tudor-SN) and alter downstream processes including cytoplasmic miRNA processing and target specificity (e. These hairpin loop structures are composed of about 70 nucleotides each. Perhaps as many as 16% of pre-miRNAs may be altered through nuclear RNA editing. [56][57][58] The RNA-induced silencing complex[edit] Main article: RNA-induced silencing complex . a transcript may serve as a pri-miRNA and a mRNA. The resulting transcript iscapped with a specially modified nucleotide at the 5’ end. Animal miRNAs are initially transcribed as part of one arm of an ∼80 nucleotide RNA stem-loop that in turn forms part of a several hundred nucleotides [42][47] long miRNA precursor termed a primary miRNA (pri-miRNA)s. DGCR8 associates with the enzyme Drosha. the site-specific modification of RNA sequences to yield products different from those encoded by their DNA. The product resulting has a two-nucleotide overhang at its 3’ end." Originally thought to exist only in Drosophila and C. although this does not necessarily mean the mature miRNAs of a family will be homologous in structure and function. are known as "Mirtrons. mirtrons have now been found in [55] mammals. The promoters mentioned have been shown to have some similarities in their motifs to promoters of other genes transcribed by RNA polymerase II such as protein [42][52] coding genes.. it has 3' hydroxyl and 5' phosphate groups. named for its association with DiGeorge Syndrome. and [53] mammalian wide interspersed repeat (MWIR) promoter units. transfer RNAs (tRNAs).with their host genes. In this complex. The double-stranded RNA structure of the hairpins in a pri-miRNA is recognized by a nuclear protein known as DiGeorge Syndrome Critical Region 8 (DGCR8 or "Pasha" in invertebrates). Other miRNA genes showing a common promoter include the 42-48% of all miRNAs originating from polycistronic units containing multiple discrete loops from which mature miRNAs [42][51] are processed. Pre-miRNAs that are spliced directly out of introns. RNA editing can halt nuclear processing (for example. elegans. of pri-miR-142. and spliced.g. bypassing the Microprocessor complex. Each hairpin is flanked by sequences necessary for efficient processing. This increases the diversity and scope of miRNA action beyond that implicated from the genome alone. RNA polymerase III(Pol III) transcribes some miRNAs. The polymerase often binds to a promoter found near the DNA sequence encoding what will become the hairpin loop of the pre-miRNA.

They bind the mature miRNA and orient it for interaction with a target mRNA. both strands of the duplex are [69] viable and become functional miRNA that target different mRNA populations. the putative DNA helicase MOV10. Members of the Argonaute (Ago) protein family are central to RISC function. for example human Ago2. argonautes may also recruit additional proteins to achieve translational [70] repression. . cleave target transcripts directly. et al. Ago2 can cleave the mRNA and lead to direct mRNA degradation. such as TNF alpha or GM-CSF (ref Jing. In some cases. uptake by the Argonaute protein is thought to stabilize the guide strand. RISC with incorporated miRNA is sometimes referred to as "miRISC. and the RNA recognition motif [59][72][73] containing protein TNRC6B. while the opposite (* or "passenger") strand is preferentially destroyed. called the passenger strand due to its lower levels in the steady state. miR16 contains a sequence complementary to the AU-rich element found in the 3'UTR of many unstable mRNAs. selected on the basis of its thermodynamic instability and [65][66][67] weaker base-pairing relative to the other strand. Q." Dicer processing of the pre-miRNA is thought to be coupled with unwinding of the duplex. During miRNA maturation in the cytoplasm. It has been demonstrated that if there is complete complementation between the miRNA and target mRNA sequence. miRNA turnover[edit] Turnover of mature miRNA is needed for rapid changes in miRNA expression profiles. [75] also known as Rat1p. Cell 2005 pmid = 15766526). if there isn't complete [20] complementation the silencing is achieved by preventing translation. only one strand is incorporated into the miRISC. The human genome encodes eight argonaute proteins divided by sequence similarities into two families: AGO (with four members present in all mammalian cells and called E1F2C/hAgo in [64][70] humans). PACT (protein activator of the interferon induced protein kinase (PACT). Tudor staphylococcal nucleasedomain-containing protein (Tudor-SN). Some argonautes. Mode of silencing[edit] Gene silencing may occur either via mRNA degradation or preventing mRNA from being translated. Yet. and PIWI (found in the germ line and hematopoietic stem cells). Similar enzymes are encoded in animal genomes. Argonautes are needed for miRNA-induced silencing and contain two conserved RNA binding domains: a PAZ domain that can bind the single stranded 3’ end of the mature miRNA and a PIWI domain that structurally res embles ribonuclease-H and functions to interact with the 5’ end of the guide strand. SDN (small RNA degrading nuclease) family members degrade miRNAs in the opposite (3'-to-5') direction. is denoted with an asterisk (*) and is normally degraded.. In what has been called a "Use it or lose it" strategy. but their [74] roles have not yet been described. leading to degradation of the non-targeting molecules.The mature miRNA is part of an active RNA-induced silencing complex (RISC) containing Dicer and many [63] [64] associated proteins. For example. the SMN complex. Generally. In plants. The other strand. fragile X mental retardation protein (FMRP). Decay of mature miRNAs in Caenorhabditis elegans is mediated by the 5´-to-3´ exoribonuclease XRN2. RISC is also known as a microRNA ribonucleoprotein complex (miRNP). The position of the stem-loop may also influence [68] strand choice. Argonaute may preferentially retain miRNAs with many targets over miRNAs with few [74] or no targets. Additional RISC components include TRBP [human immunodeficiency virus (HIV) transactivating [71] response RNA (TAR) binding protein].

An extra A added to the end of mammalian miR-122. and miRNA serves as a template for recognizing specific mRNA sequences. M8) on the degradation of mRNA. mature plant miRNAs appear to be stabilized by the addition of methyl moieties at the 3' end.Several miRNA modifications affect miRNA stability. The function of miRNAs appears to be in gene regulation. ribosome drop-off. The actual work of RNA silencing is performed by RISC (RNA-induced silencing complex) in which the main catalytic subunit is one of the Argonaute proteins (AGO). M7. 80S is the assembled ribosome bound to mRNA. For that purpose. a miRNA is complementary to a part of one or moremessenger RNAs (mRNAs). M2) on the ribosome assembly. co-translational protein degradation and others) that are not visualized here. The initiation of mRNA can proceed in a cap-independent manner. transport to P-bodies. [76] Here. There exist other mechanisms of microRNA action on protein translation (transcriptional. M3) on the translation process. a liver-enriched miRNA important in Hepatitis C. Animal miRNAs are usually complementary to a site in the 3' UTR whereas plant miRNAs are usually complementary to coding . Cellular functions[edit] Interaction of microRNA with protein translation process. stabilizes the molecule. Both plant and animal miRNAs may be altered by addition of adenine (A) residues to the 3' end of the miRNA. eIF4F is an translation initiation factor. the consequences of this modification are incompletely understood. As indicated by work in the model organism Arabidopsis thaliana (thale cress). a modification that may be associated with miRNA degradation. PABC1 is the Poly-A binding protein. and "cap" is the mRNA cap structure needed for mRNA circularization (which can be the normal m7G-cap or artificial modified A-cap). through recruiting 40S to IRES (Internal Ribosome Entry Site) located in 5’UTR region. Several (from nine documented) mechanisms of translation repression are shown: M1) on the initiation process. However. and [74] plant miRNAs ending with an adenine residue have slower decay rates. preventing assembling of the initiation complex or recruiting the 40S ribosomal subunit. 40S and 60S are light and heavy components of the ribosome. The 2'-O-conjugated methyl groups block the addition of uracil (U) residues by uridyltransferase enzymes. Uridylation of some animal miRNAs has also been reported. uridylation may also protect some miRNAs.

nucleotides 2 –7 of the miRNA (its 'seed [6][16] [80] region' ) still have to be perfectly complementary. Recent work on miR-430 shows that translational repression is caused by the [83] disruption of translation initiation. genes involved in functions common to all cells. Interactions between microRNAs and complementary sequences on genes and even pseudogenes that share sequence homology are thought to be a back channel of communication regulating expression levels between paralogous genes. For partially complementary microRNAs to recognise their targets. Sequestration in P-bodies.regions of mRNAs. 9. Cap-40S initiation inhibition. 4. or a combination of the two is hotly debated. 3. miRNAs occasionally also cause histone modification and DNA methylation of promoter sites. Co-translational nascent protein degradation. The match-ups are imperfect. However. mRNA Decay (destabilisation). Nine mechanisms of miRNA action are described and assembled in a unified mathematical model: 1. Nevertheless. these microRNAs bind to "microRNA response elements" on genes and pseudogenes and may provide another [89] explanation for the persistence of non-coding DNA. MicroRNAs that are partially complementary to a target can also speed up deadenylation. Transcriptional inhibition through microRNA-mediated chromatin reorganization following by gene silencing. This was demonstrated in human cells using synthetic dsRNAs termed small activating [87] [88] RNAs (saRNAs). a mechanism that has been termed "small RNA-induced gene activation" or RNAa. It is often impossible to discern these mechanisms using the experimental data about stationary reaction [76] rates. 60S Ribosomal unit joining inhibition. While degradation of miRNA-targeted mRNA is well documented. 5. . causing mRNAs to be degraded [82] sooner. which [84][85] affects the expression of target genes. 8. the animal microRNAs target a diverse set of genes. whether or not translational repression is accomplished through mRNA degradation. Unlike plant microRNAs. they are differentiated in dynamics and have different kinetic signatures. but has also been demonstrated for endogenous microRNA. 2. Perfect or near perfect base pairing with the target RNA promotes cleavage of the [78] [79] RNA. have relatively fewer microRNA target sites and seem to be under selection to avoid targeting by microRNAs. Ribosome drop-off (premature termination). This is the primary mode of plant miRNAs. mRNA Cleavage. such as gene expression. Animal miRNAs inhibit protein translation of the [81] [79] target mRNA (this exists in plants as well but is less common). dsRNAs targeting gene promoters can induce potent transcriptional activation of associated genes. Elongation inhibition. [86] [16] [76] [77] dsRNA can also activate gene expression. independent of mRNA deadenylation. In animals miRNAs more often have only partly the right sequence of nucleotides to bond with the target mRNA. Given the name "competing endogenous RNAs" (ceRNAs). 7. 6. translational inhibition.

MicroRNAs can also form from inverted duplications of protein-coding sequences. Their origin may have permitted the development of morphological innovation. but also by the [94] duplication and modification of existing microRNAs. The activity of an miRNA can be experimentally inhibited using a locked . the difference in how these microRNAs function and the way they are processed suggests that [100] microRNAs arose independently in plants and animals. partly due to their length. This makes them a valuable phylogenetic marker. older microRNAs have a much lower rate of change (often less than one substitution per hundred million [93] years). Variations of this method achieve absolute or relative [105] quantification.Evolution[edit] MicroRNAs are significant phylogenetic markers because of their astonishingly low rate of [90] evolution. and they are being looked upon as a possible [98] solution to such outstanding phylogenetic problems as the relationships of arthropods. miRNAs can also be hybridized to microarrays. from the brown algae to the animals. New microRNAs are created in multiple different ways.e. so that relative levels of miRNAs can be determined in different [106] samples. Indeed. In Arabidopsis thaliana.2 and 3. The rate of evolution (i. however. However. nucleotide substitution) in recently originated microRNAs is comparable to that elsewhere in the non-coding DNA.3 genes per million [97] years. ultimately. although microRNAs [94] that are more recently derived (and thus presumably non-functional) are frequently lost. [99] Experimental detection and manipulation of miRNA[edit] While researchers have focused on the study of miRNA expression in physiological and pathological processes. MicroRNAs are degraded much more easily than mRNAs. suggesting that once a microRNA gains a function it undergoes extreme purifying [94] selection. the net flux of miRNA genes has been predicted to be between 1. This makes it necessary to cool samples on ice and use RNase-free equipment whenever working with [104] microRNAs. complex [93] life. Whilst short RNA sequences (50 – hundreds of base pairs) of a [102] broadly comparable function occur in bacteria. a microRNA is rarely lost from an animal's genome. The stability of the [103] stored miRNA samples has often been questioned. At this point. different regions within an miRNA gene seem to be under different evolutionary pressures. Additionally. implying evolution by neutral drift. MicroRNA expression can be quantified in a two-step polymerase chain reaction process of modified RTPCR followed by quantitative PCR. Across all species. Novel microRNAs can originate from the random formation of hairpins in "non-coding" sections of DNA (i. in excess of 5000 had [101] been identified by March 2010.e. various technical variables related to microRNA isolation have emerged. permitted the genesis of complex organs and perhaps. MicroRNAs feature in the genomes of most eukaryotic organisms. MicroRNAs can be both discovered and profiled by high-throughput sequencing methods [107] (MicroRNA Sequencing). rapid bursts of morphological innovation are generally associated with a high rate of [90][92] microRNA accumulation. and by making gene expression more [92] specific and 'fine-tunable'. MicroRNAs origin as a regulatory mechanism developed from previous RNAi machinery [91] which was initially used as a defense against exogenous genetic material such as viruses. slides or chips with probes to hundreds or thousands of miRNA targets. bacteria lack true microRNAs. but also because of the ubiquitously present RNases. introns or intergene regions). where regions that are vital for processing and function have much higher levels of [96] [93] conservation. which allows for the creation of a foldback hairpin [95] structure.

and miRNA-expression information have been [120][121] proposed. ideas for analysis tools that integrate mRNA. making it ideal for detection of short miRNA. To pair mRNA. [123] documents known relationships between miRNA dysregulation and human disease. in ). a Morpholino oligo or a 2'-O-methyl RNA oligo. publicly available database. A manually curated. For the ―in situ‖ detection of miRNA. [124] miRNA and cancer[edit] Role of miRNA in a cancer cell The first human disease known to be associated with miRNA deregulation was chronic lymphocytic [122] leukemia and later many miRNAs have been found to have links with some types .and miRNA[118][119] data. The locked conformation of LNA results in enhanced hybridization properties and increases [115] sensitivity and selectivity. MicroRNA maturation can be inhibited at several [111] points by steric-blocking oligos.miR2Disease. mRNA-expression is therefore often [116][117] analyzed as well to check for miRNA-effects in their levels (e. High-throughput quantification of miRNAs is often difficult and prone to errors.nucleic acid (LNA) oligo. g. LNA or Morpholino probes can be used. g. A mutation in the [125] seed region of miR-184 causes hereditary keratoconus with anterior polar cataract. a specific miRNA can be silenced by a complementary antagomir. because of high expression levels). Additionally. [108][109] [110] miRNA and disease[edit] Just as miRNA is involved in the normal functioning of eukaryotic cells. for the larger variance (compared to mRNAs) that comes with the methodological problems. miRNA and inherited diseases[edit] A mutation in the seed region of miR-96 causes hereditary progressive hearing loss. The miRNA target site of an mRNA transcript can also be blocked by a [112] [113] [114] steric-blocking oligo. While this is usually done after miRNAs of interest have been detected (e. databases can be used which predict miRNA-targets based on their base sequence. Deletion of the [126] miR-17~92 cluster causes skeletal and growth defects. so has dysregulation of miRNA [122] been associated with disease.

Sufficient selectivity and specificity could be achieved using small (less than 1 mL) samples of blood. A further advantage is they can also be performed at each consultation to assess disease response and detection of early relapse. non-invasive [135][136] way to identify at-risk patients who should undergo colonoscopy. for example one study on NSCLC samples found that low miR-324a levels could serve as a prognostic indicator of poor [137] survival. miR-494. are overexpressed in cancer. and miR-1973 are promising disease response [139] biomarkers. Those with a rapid response to initial treatment may benefit from truncated treatment regimens. Cell-free miRNA are highly stable in blood. Another study found that two types of miRNA inhibit the E2F1 protein. In classical Hodgkin Lymphoma. miRNA appears to bind to messenger RNA before it can be translated to proteins that switch genes on [131] and off. Leukemia can be caused by the insertion of a viral genome next to the 17-92 array of microRNAs leading to increased [130] expression of this microRNA. and are quantifiable within the diagnostic laboratory. resectable (Stage II) colorectal cancer could be distinguished from those of sex-and age-matched healthy volunteers. By measuring activity among 217 genes encoding miRNA. Early results showed that blood plasma samples collected from patients with early. [122][127][128] . Mice that were engineered to produce a surplus of types of miRNA found in lymphoma cells developed the disease within 50 days and [127] died two weeks later. Another role for miRNA in cancers is to use their expression level as a prognostic. A novel miRNA-profiling based screening assay for the detection of early-stage colorectal cancer has been developed and is currently in clinical trials. This will allow doctors to determine the original tissue type which spawned a cancer and to be able to target a treatment [132] course based on the original tissue type. MicroRNA-21 is involved in several cancer-types such as glioblastoma and astrocytoma was one of the first microRNAs to be identified as an [129] oncomir. patterns of gene activity that can distinguish types of cancers can be discerned. Circulating miRNAs have the potential to greatly assist clinical decision making and aid interpretation of positron emission tomography combined with computerized tomography. A study of mice altered to produce excess c-Myc — a protein with mutated forms implicated in several cancers — shows that miRNA has an effect on the development of cancer. miRNA signatures may enable classification of cancer.of cancer and are sometimes referred to as "oncomirs". thus the need for more accurate measures of disease response. Transgenic mice that over-express or lack specific miRNAs have provided insight into the role of small [133] RNAs in various malignancies. mice without the surplus miRNA lived over 100 days. another found that either high miR-185 or low miR-133b levels correlated with metastasis and [138] poor survival in colorectal cancer. In contrast. Optimal treatment for cancer involves accurately identifying patients for risk-stratified therapy. Much work has also been done on the role of microRNAs in establishing and maintaining cancer stem cells that are especially resistant to chemotherapy and often [134] responsible for relapse. which regulates cell proliferation. plasma miR-21. The test has potential to be a cost-effective. miRNA profiling has already been able to determine whether [128] patients with chronic lymphocytic leukemia had slow growing or aggressive forms of the cancer.

damage tends to accumulate in DNA. and expression of these proteins is regulated by microRNAs. Accumulated DNA damage can also cause epigenetic alterations due to errors during DNA [143][144] repair. HMGA2 protein specifically targets the promoter of ERCC1. miR-16. If repair is deficient. in normal tissues. which reduces protein expression of MGMT. Such DNA damage can cause mutational errors during DNA replication due to error-prone translesion synthesis. These proteins have three highly positively-charged regions. ERCC1 protein expression was deficient in 100% of 47 evaluated colon cancers (though the [154] [155] extent to which HGMA2 was involved is not known). termed AT hooks. DNA repair and cancer[edit] Individuals with an inherited deficiency in DNA repair capability are at increased risk of cancer (also see DNA repair-deficiency disorder). including thyroid. However. Human neoplasias. showed that HMGA1 protein binds to the promoter region of DNA repair gene BRCA1 and inhibits BRCA1 promoter activity. Five members of the microRNA-200 family (miR-200a. which represses MLH1 expression. HMGA1b and HMGA2) are implicated in cancer. Germ line mutations in DNA repair genes cause only 2–5% of colon cancer cases. prostatic. increased expression of miR181d and reduced expression of DNA repair enzyme MGMT may be a causal factor.. They also showed that while only 11% of breast tumors had hypermethylation of the BRCA1 gene. 82% of aggressive breast cancers have low BRCA1 protein expression. in 28% of glioblastomas. up to 15% of the MLH1-deficiencies in sporadic colon cancers appeared to be due to [147] over-expression of the microRNA miR-155. However. HGMA proteins are polypeptides of ~100 amino acid residues characterized by a modular sequence organization. HMGA proteins (HMGA1a. colorectal. miR-200c. [140] miRNA. Among 68 sporadic colon cancers with reduced expression of the DNA mismatch repair protein MLH1. causing DNA repair deficiencies. Palmieri et al. Transgenic mice with HMGA1 targeted to lymphoid cells develop aggressive lymphoma. that bind the minor groove of AT-rich DNA stretches in specific regions of DNA. cervical. Baldassarre et al. thus reducing expression of this DNA repair [153] gene. but that the HMGA1 gene can act as an oncogene to cause [151] [152] cancer. However.Recent studies have miR-205 targeted for inhibiting the metastatic nature of breast cancer. miR-200b. showed that. most were found to be deficient due to epigenetic methylation of the CpG island of the MLH1 [146] gene. HGMA1 and HMGA2 genes are targeted (and thus strongly reduced in expression) by miR-15. In 29–66% of glioblastomas. DNA repair is deficient due to epigenetic methylation of the MGMT gene. In the glioblastomas without methylated MGMT promoters. pancreatic and ovarian [150] carcinoma. are frequently associated with cancers and may be an important causal factor for these cancers. [148][149] [145] [142] . Thus. the [148] MGMT protein is deficient but the MGMT promoter is not methylated. altered expression of microRNAs. show a strong increase of HMGA1a and HMGA1b proteins. and most of these reductions were due to chromatin remodeling by high levels of HMGA1 protein. showing that high HMGA1 expression is not only associated with cancers. Such mutations and epigenetic alterations can give rise to cancer (see malignant neoplasms). for 28% of glioblastomas. HMGA expression is almost undetectable in differentiated adult tissues but is elevated in many cancers. miR-141 and miR-429) are down [141] regulated in tumour progression of breast cancer. the level of microRNA miR-181d is inversely correlated with protein expression of MGMT and the direct target of miR-181d is the MGMT mRNA 3’UTR (the three prime [148] untranslated region of MGMT mRNA).

and 222 significantly inhibited adipogenesis and repressed induction of the master regulators PPARγ and CCAAT/enhancer-binding protein alpha [177] (CEBPA). For two of these. BRCA1 (miR-182) and P53 (miR-504. MSH2 (miR-21). . miRNA expression profiling studies demonstrate that expression levels of specific miRNAs change in diseased human hearts. This paves the way for possible obesity treatments on the genetic level. suggesting that they act as negative regulators of differentiation.221.synapse formation and synapse maturation (where miR-134 and miR-138 are thought to be [172] [173][174] involved). miR[157] 182. miR-15 and miR-16. then HMGA1 and HMGA2 proteins are expressed at a high level. miR-196a and Let[156][157] 7a) have methylated promoters and therefore low expression in colon cancer. miRNA and the nervous system[edit] miRNAs appear to regulate the nervous system. Wan et al. In contrast to the previous example. Some studies find altered miRNA expression in schizophrenia. when compared with the normal pituitary gland. where under-expression of miRNAs indirectly caused reduced expression of DNA repair genes. [171] miRNA and obesity[edit] miRNAs play crucial roles in the regulation of stem cell progenitors differentiating [175] into adipocytes. miRNA and heart disease[edit] The global role of miRNA function in the heart has been addressed by conditionally inhibiting miRNA maturation in the murine heart. including the regulation of key factors important for cardiogenesis. Studies to determine what role pluripotent stem cells play in adipogenesis. Consistent with the down-regulation of these HMGA-targeting miRNAs. Three of these microRNAs (miR-16. including dendritogenesis (involving miR-132.miR-221. in some cases over-expression of certain miRNAs may directly reduce [160] expression of specific DNA repair proteins. Thus DNA repair can be reduced. Conversely. the hypertrophic growth response. HMGA1 and HMGA2 target (reduce expression of) BRCA1 and ERCC1 DNA [159] repair genes.miR-26a. miR-373). and has revealed that miRNAs play an essential role during its [161][162] development. miR-125b). referred to 6 DNA repair genes that are directly targeted by the miRNAs indicated: ATM (miR-421). Three of these miRNAs (miR-21. RAD52 (miR-210. studies on specific miRNAs in animal models have identified distinct roles for miRNAs both during heart development and under pathological conditions. When these microRNAs are expressed at a low level. Neural miRNAs are involved at various stages of synaptic development. miR-196a2 and Let-7a. Decreased expression of miR-155. have been found during the adipogenic programming of both immortalized and primary hMSCs. However. an increase in the HMGA1 and HMGA2-specific mRNAs was observed. RAD23B (miR-373). miR-125b) are among those identified by Schnekenburger and Diederich as over-expressed in colon cancer through epigenetic hypomethylation. Furthermore. pointing to their involvement [163][164][165] incardiomyopathies. and cardiac [162][166][167][168][169][170] conductance.and miR-222. ectopic expression of the miRNAs 155. each of these HMGA-targeting miRNAs are drastically reduced in almost all human pituitary adenomas studied. were [176] examined in the immortalized human bone marrow-derived stromal cell line hMSC-Tert20. miR-134 and miR124). Over expression of any one of these miRNAs can cause reduced expression of its target DNA repair gene. likely contributing to cancer progression. the coding regions are epigenetically silenced in cancer due to histone deacetylase [158] activity.

Either of the strands becomes stably associated with RNA-induced silenced . or the pre-miRNA. The sequential processing reaction excises the stem-loop from the remainder of the transcript to create a pre-miRNA product. and remarkably resistant to high fat dietinduced obesity and diabetes. Figure 1. Then this pre-miRNA is actively transported from the nucleus to the cytoplasm by Ran-GTP and export receptor. One end of the mature miRNA was cut by Drosha in nuclear and the other end is processed in the cytoplasm by the enzyme Dicer. is the let-7 family. which occurs in the nucleus and is mostly carried out by a nuclear member of the RNase III family (Drosha). These experimental findings suggest that let-7 inhibition could represent a new therapy for obesity and type 2 diabetes. which can be called miRISC complex[15. and thus more prone to high [178] fat diet-induced obesity and diabetes. miRNA and non-coding RNAs[edit] When the human genome project mapped its first chromosome in 1999. miRNA and viruses[edit] The expression of transcription activators by human herpesvirus-6 DNA is believed to be regulated by [185] viral miRNA. However. which is carried out by the canonical Dicer enzyme in the cytoplasm. 2004). with transcripts capped and polyadenylated. only around 20. it was predicted the genome would contain over 100. The miRISC complex acts as a regulator of target gene by specially recognizing and regulating particular mRNAs to inhibit target genes [17]. and diabetes. In contrast when let-7 was inhibited by injections of let-7specific antagomirs. including several snoRNAs and miRNAs. mice became insulin-resistant. and experimental [183] validation (including the creation of miRNA derived antisense oligonucleotides called antagomirs) have [184] revealed that many transcripts are non protein-coding RNA. the advent of bioinformatics approaches combined with genome tiling studies examining the [181] [182] transcriptome. Biological functions of miRNA. Their transcription is typically performed by RNA polymerase II. Either of the strands becomes stably associated with RNA-induced silenced complex (RISC).14] and are encoded by a bewildering array of genes. When let-7 was ectopically overexpressed to mimic accelerated aging. it could also [179] reverse and cure diabetes.16]. MiRNAs involved in cancer cell metabolism The biogenesis of miRNAs is tightly associated with their action mechanism (Figure 1). Let-7 is known to accumulate in human tissues during the course of aging. Not only could let-7 inhibition prevent obesity and diabetes. The first step is the nuclear cleavage of the pri-miRNA. systematic sequencing of full length cDNA libraries. known as the miRNA precursor. mice become more insulin-sensitive. obesity. The following step excises the terminal loop from the pre-miRNA stem to create a mature miRNA duplex of approximately 22 bp length.000 were eventually [180] identified (International Human Genome Sequencing Consortium. Most miRNAs derived from independent transcription units [13.000 protein coding genes. Since then. with a ~60-70 nt stem loop intermediate liberated. The resulting primary or pri-miRNA transcript extends both 5’ and 3’ from the miRNA sequence.Another class of miRNAs that regulate insulin resistance.

In addition. Metabolic shift in cancer cells seems to be influenced by oncogene and tumor suppressor networks [22]. miR-422.complex (RISC). including participating in controlling Acetyl-CoA and plasma cholesterol. The functional association analysis of miRNAs and metabolic pathways uncovered that miRNAs predominantly regulate central metabolic pathways such as amino acid biosynthesis. miR-506 and miR-136. ( C) inducing ribosomes to drop off prematurely thus repressing the translation initiation and (D) promoting mRNA degradation. miR-27a. miR-34a. Tibiche and Wang systematically analyzed the human metabolic network by integrating miRNA target genes into the network [23]. A shift in glucose metabolism from oxidative phosphorylation to aerobic glycolysis was a key biochemical hallmark of tumor cells [18. a lipid kinase that regulates the levels of phosphorylated phosphatidylinositol at the plasma membrane. . which consists of an increase in glycolysis maintained in conditions of high oxygen tension and gives rise to enhanced lactate production [20. which can be called miRISC complex. including p53. miR-138. certain sugar and lipid metabolism (Figure 2). lipid and amino acid. miR-122 and miR-33a/b function on lipid metabolism. And miR-14. In addition. Figure 2. What’s more. MiRNAs could directly modulate the expression of metabolic transporters or enzyme activities. which is targeted by miR-320. more than 20 miRNAs. The main miRNAs involved in metabolism of glucose. miR-25. Among the total 60 miRNAs mentioned in the text.21]. They performed randomization tests to determine whether a multiple-gene-node is significantly regulated by miRNAs and defined 79 multiple-gene-nodes as miRNA targets. including directly targeting key molecules (transporters or enzymes / kinases) of metabolic processes and regulating multiple oncogenic signaling pathways (Figure 3).) play functions in amino acid metabolism mainly through regulating acyltransferase and α-ketoacid dehydrogenase. While several miRNAs (miR-23b*.. miR-277 etc. The miRISC complex inhibits the target genes by (A) repressing initiation at the cap recognition. AMPK and AKT signaling pathway. miR-29a/b. miR-370.) participate in metabolism-associated oncogenic signaling pathways. (B) inducing deadenylation of mRNA and thereby inhibiting circularization of mRNA. miR123a. miR-504. plays a key role in cancer cell metabolism. There are several lines of evidence that many key molecules in cell metabolism are miRNA targets. MiRNAs also play pivotal roles in the expression level of transcription factors and oncogenes or tumor suppressors. They merged the miRNA targets of single-gene-nodes with the multiple-gene-nodes. Regulation of metabolic activity by miRNAs MiRNAs regulate cell metabolic processes through complicated mechanisms.19]. about 29% of the mentioned miRNAs (miR-125b. thus giving a clue that miRNA regulates cell metabolism. etc. involve in glucose metabolism. most of these tumor suppressors are miRNA targets. and found that 238 (22%) nodes are miRNA targets. etc. The altered metabolism was called “Warburg phenomenon”. Since miRNAs regulate a substantial fraction of genes in animal genomes. c-Myc. miR-133. miR-146. miR-199a. miR-335. phosphatidylinositol 3-kinase. miR30d. as well as metabolismassociated oncogenic signaling pathways . including miR375. For example.

c-Myc and AKT/mTOR signaling pathways. particularly estrogen. In the initial step of glucose metabolism. The regulation of several specifically signaling pathways involved in cancer cell metabolism by miRNAs will be introduced in detail in the second section of this review. To date. phosphatidylinositol 3kinase. And the specific miRNAs in glucose metabolism will be summarized into 4 subtitles as follows. including miRNA effects on glucose uptake. for example. A study in renal cell carcinoma . glycolysis.6-bisphosphate is broken down into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. aerobic glycolysis in tumor cells is driven by multiple miRNA-involved oncogenic signaling pathways. Several factors have been implicated in the regulation of their expressions. high glucose requirements. The potential effects of the GLUTs level seem to facilitate accelerated metabolism. mRNA dysregulation in any step of metabolic processes contributes to metabolic abnormalities and even cancer development. In addition. is regulated by miR21 [28]. HIF. AKT. glucose could be transported over a plasma membrane by GLUT3 or GLUT4 which is a target of miR-133 [24] or miR-195-5p [25]. For example.MiRNAs could regulate cell metabolism by modulating the expression of metabolic transporters (like GLUT) or enzymes (HK2. the first rate-limiting enzyme of glycolysis. The amounts of the GLUT1. lactate dehydrogenase isoform A. GLUT. and increased glucose uptake in malignant cells. pyruvate dehydrogenase. The steps regulated by miRNAs are indicated by red circular arrows. LAT1. FASN. a cardinal node in diverse signaling cascades. The molecular mechanisms driving the Warburg effect in cancer cells were taken as an example to explain miRNA regulation in energy metabolism. hypoxic also drives GLUT expression [31] as well as metabolic-stress-induced signaling pathways. and GLUT3 transcripts were elevated in most cancer tissues. L-type amino acid transporter 1. which stimulates glycolysis by directly regulating glycolytic enzymes and activating downstream mammalian target of rapamycin (mTOR) activity. PDK. ALDOA and PDK1) and acting on p53. the hexokinase 2 (HK2). fructose 1.Figure 3. While the enzyme Aldo A is down-regulated by miR-15a/16-1cluster [27]. As shown in Figure 3. Hormonal. MiRNAs could regulate glucose uptake via altering the GLUTs expressions. while mRNA levels of GLUT4 and GLUT5 were below sensitivity in these cancer tissues. several miRNAs affect gene transcription and expression of glucose transporters (GLUTs) which are responsible for transporting glucose into cytoplasm. GLUT2. Therefore. could provide a mechanism of GLUT regulation [30]. glucose transporter. PDH. such as adenosine monophosphate-activated protein kinase (AMPK). 14 members of GLUTs have been identified [29]. On the other hand. MicroRNAs regulate cell metabolism by targeting key metabolic enzymes and multiple oncogenic signaling pathways. fatty acid synthase. ovarian hormones. LDH-A. hypoxia-inducible factor. pyruvate dehydrogenase kinase. triggering upregulation of GLUT receptors [32]. MCT. PI3K. MiR-133 has been confirmed to regulate the expression of GLUT4 by targeting KLF15 in a rat model [31]. In the following step. tricarboxylic acid (TCA) cycle and insulin regulation. is among the top list of genes predicted and potentially regulated by multiple miRNAs including miR-143 [26]. which is catalyzed by aldolase A (Aldo A) in the reversible aldol reaction. Thus. MiRNAs regulate glucose metabolism MiRNAs affect glucose uptake GLUTs (or SLC2A) are a wide group of membrane proteins that facilitate the transport of glucose over a plasma membrane in most mammalian cells. monocarboxylate transporter. and the related miRNAs are listed in the bracket. Along the glycolysis reaction chain. miRNAs could directly regulate intracellular glucose levels.

miR-19a and miR-133a are altered in colorectal carcinoma [34]. a transcriptional activator for HK2 [35]. miR-130b is highly down-regulated in pancreatic tumors. miR-143. These reports all illustrated that miR-143 targets HK2 to regulate glucose metabolism in cancer cells. There have been over 200 publications on circulating miRNA in cancers including prostate. as an essential regulator of glycolysis. Considering the sources of variation. The knowledge that miRNA expression is frequently dysregulated in cancer has uncovered an entirely new repertoire of molecular factors upstream of gene expression. since they are abundant in blood. miR-138.demonstrated that down-regulated miR-199a. MiRNAs as biomarkers for human cancer By targeting and controlling the expression of mRNA. tissue specificity. the reduced levels of miR-126. and it is a potential cancer therapeutic target. and prostate)[96]. and the miR-15a/16-1 cluster could reduce the levels of Aldo A [27]. whereas an increased expression of miR-130b. In addition. there has been an accumulating body of evidence to support circulating miRNAs as non-invasive. metastatic potential. Interestingly. except by activating the signal transducer and activator of transcription 3. HK2 has been validated as a miR-143 target and thus miR-143 could affect glucose metabolism in colon cancer cells [37]. In this process. inflammation-related miR-155. On the other hand. ease of detection and manipulation. miRNAs can control highly complex signal transduction pathways and multiple metabolic processes. miRNAs also regulate other important intermediate steps in the glycolysis pathway. present their potential as tumor markers [97]. state of microRNA in plasma and origin and implications . pancreatic. What’s more. and their roles in regulating GLUT expression might explain the disordered metabolism in colorectal carcinoma. colon. Besides. sensitive biomarkers of disease states. particularly cancers (breast. For example. lung. Exploiting the unique characteristics of these molecules including their stability. and other clinical characteristics for several types of cancers. miR-155 could repress miR-143 thereby upregulating the expression of HK2 at the post-transcriptional level. Functions of miRNAs on glycolysis Studies show that miRNAs regulate the irreversible steps in glycolysis. modulates glycolysis via targeting HK2 [12]. miR-9 and miR-9* (expressed from the 3' mature sequence). In addition. The tissue concentrations of specific miRNAs have been associated with tumor invasiveness. members of the miR-17-92 cluster. highly stable and disease associated. miR-143 has also been identified as an essential regulator of cancer glycolysis via targeting HK2 in human lung cancer [12]. colorectal. thus committing glucose to the glycolytic pathway. miR-19b and miR301a can result in the down-regulation of GLUT-1 [32]. Thus miR-122 and miR-15a/16-1 cluster are involved in glycolysis in cancer cells. The oxysterol-binding-protein-related-protein 8 has been revealed as a target of miR-143 by quantitative mass spectrometry analysis [34]. which phosphorylates glucose to produce glucose 6-phosphate. For example. Recently new protein targets of miRNAs have been identified by sensitive mass spectrometric studies. the above articles were published almost at the same time. miR-150 and miR-532-5p were correlated with an increased expression of GLUT-1. will bring clinicians ever closer to achieving the goal of individualized cancer treatment [94]. with exciting potential as novel biomarkers and therapeutic targets in cancer [93]. and smooth muscle-enriched miR-145 in patients with coronary artery disease compared with healthy controls [98]. hepatic. For example. Furthermore. which are usually involved in different oncogenic pathways [92]. What’s more. lung. miR-143 inhibits the expression of HK2 both in primary keratinocytes and in head and neck squamous cell carcinoma-derived cell lines [36]. breast. Likewise.6-bisphosphate is broken down into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. ovarian. miR-122 was predicted to target Aldo A [26. and changes in miRNA within a tissue type can be correlated with disease status. miR-19a. serum microRNAs are attractive disease biomarkers [100]. lung. miRNAs are produced in a tissue-specific manner. pancreas. Interestingly. and breast. MiR-195-5p has been identified as a direct regulator of GLUT3 by targeting GLUT3 3’-untranslated region in bladder cancer T24 cells [33]. On the one hand. including chronic lymphocytic leukemia. ovarian and leukemia since 2008.38]. especially the key enzymes[33]. published data showed that plasma miR-29a and miR-92a have strong potential as novel noninvasive biomarkers for early detection of colorectal carcinoma [99]. mostly neuronal and thus expressed in central nervous system tumors but absent in other tumors. Except for targeting the irreversible rate-limiting steps. The enzyme Aldo A catalyzes a reversible aldol reaction in which fructose 1. and prostate cancers [95]. easy to measure. and its role in regulating GLUT-1 expression might explain the increased glucose uptake in pancreatic adenocarcinoma [35].

endothelial miR-126 is deregulated in patients with type 2 diabetes. miRNA expression profiling. which may ultimately lead to novel biomarkers for risk estimation and classification and could be exploited for miRNA-based therapeutic interventions of vascular complications associated with this disease [107]. solution hybridization detection etc. A significantly increased or decreased alteration in the level of the miRNA gene product in the detected sample is indicative of the subject either having or being at risk for developing a cancer. So far. Nevertheless. data analysis of miRNA expression level and cancer risk assessment. This profile could be hopefully used for early detection of cancer. quantitative or semi-quantitative RT-PCR. Since these challenges lie on the way. For example. In addition. which in turn regulate metabolic enzymes. The level of the at least one miRNA gene product can be measured using a variety of techniques (microRNA chip. miRNAs could mainly have two ways to regulate cellular metabolism. Figure 4. thus lighting up one aspect of miRNA therapeutics. miRNAs could regulate the production of certain metabolites by directly regulating the genes that encode metabolic enzymes [101].for disease specificity. 1 Eric A. miRNAs could regulate mRNAs through chromatin remodeling [102]. miRNA expression profiles of potential patients could be assessed by measuring circulating miRNAs in patient serum. Alternatively. In general. The workflow of detecting cancer based on miRNA profiling is included sample (serum or tissue) collection. a new class of miRNA-based drugs that are capable of targeting molecules outside the range of traditional medicinal chemistry. The use of miRNAs. Moreover. Additionally. physiology and disease. such as oligonucleotide complementary[103] or antisense oligonucleotides [104] in miRNA inhibition. The emergence of miRNAs as important regulators of metabolism has garnered much interest not only from a scientific point of view but also from a clinical perspective. Abstract Go to: .106]. MiRNA-based diagnosis strategies for cancer. their clinical implementation will require improvements in drug composition and delivery. Conclusion and perspective MiRNAs are important regulators of numerous aspects of metabolic homeostasis. cost-effective miRNA profiling strategies and larger studies are needed to determine its advantage for cancer diagnosis. Miska. to suppress cell metabolism altering will hopefully lead to a new therapeutic strategy for malignant cancer [105. Because miRNAs can also regulate other non-coding RNAs. drug target identification and clinical treatment in the future (Figure 4). The function of miRNAs on cellular metabolism reveals molecular strategies for controlling metabolic flux by miRNAs in living organisms. The level of at least one miRNA gene product in a test sample from the subject is compared to that in a control sample. a variety of new strategies to identify and characterize the targets of individual miRNAs have been developed. Stefanie Sassen. the successful development of miRNA biology technologies could ultimately translate our understanding of miRNA functions in cancer into strategies for the control of cancer.2 and Carlos Caldas1 Author information ► Article notes ► Copyright and License information ► This article has been cited by other articles in PMC. molecular strategies for cancer therapy by miRNAs are still in their infancy. MiRNAs could regulate transcription factors or signaling proteins. these interactions will increase the complexity of gene regulation. MiRNAs are promising in the diagnosis of cancer. northern blot analysis.) to provide a profile for the test sample.

This phenomenon was called RNA interference (RNAi). ribosomal RNA (rRNA). The encoding DNA sequence is often referred to as an RNA gene. functional or non-coding RNA molecules are transcribed from a DNA sequence. preventing protein synthesis. the mechanism of RNA interference (RNAi) was discovered at that time. Surprisingly. It was then shown that RNA interference was mediated by 22 nucleotide single-stranded RNAs termed small interfering RNAs (siRNAs) derived from the longer double-stranded RNA precursors [87]. The small interfering RNAs were found to repress genes by eliminating the corresponding messenger RNA transcripts. The conservation of let-7 across species suggested an important and fundamental biological role for this small RNA. Seven years later. [70] discovered a second 22-nucleotide small RNA of this type. In contrast. Functional RNA genes in the human genome include transfer RNA (tRNA). more than 100 additional small regulatory RNAs similar to lin- . The lin-4 and let-7 small regulatory RNAs soon became very exciting for two reasons. Over the following years.The “small RNA revolution” Small ribonucleic acid (RNA) can act as a specific regulator of gene expression. Precursors of these miRNA molecules form structures of double-stranded RNA that can activate the RNA interference machinery. and it became clear that miRNA and RNAi pathways were intricately linked and shared common components. let-7. The first miRNA was discovered in 1993 by Victor Ambros and colleagues Rosalind Lee and Rhonda Feinbaum [42]. RNA is usually thought of as messenger RNA that serves as the template for translation of genes into proteins. This discovery has been an exciting breakthrough in Biological Sciences of the past decade. a millimeter-long animal used as a model organism in biological research. homologs of the let-7 gene were identified in other animals including humans [65]. Building on previous work mainly in plants [50]. [23] discovered that exogenous double-stranded RNA can be used to specifically interfere with gene function. termed lin-4. identified genes involved in developmental timing [42]. Fire et al. did not encode a protein but instead a novel 22-nucleotide small RNA. culminating in last year’s Nobel Prize in Physiology or Medicine awarded to Andrew Fire and Craig Mello. A genetic screen in the roundworm Caenorhabditis elegans. and thus. many new small functional RNAs have been found. Within the following year. MicroRNAs downregulate gene expression either by degradation of messenger RNA through the RNA interference pathway or by inhibiting protein translation. one of the genes. a gene also involved in C. and various other small non-coding RNAs. They also speculated that organisms might use double-stranded RNA naturally as a way of silencing genes. Reinhart et al. Firstly. Secondly. Several hundred genes in our genome encode small functional RNA molecules collectively called microRNAs (miRNAs). but not translated into protein. elegans developmental timing.

the . flies. Two ribonuclease enzymes.. In humans. 41. The ribonuclease Drosha excises the stem-loop structure to form the precursor miRNA (or pre-miRNA) [43]. termed miRNA*. In the nucleus. 1b)... The mature miRNA is partially complementary to one or more messenger RNAs. 40. 40. 48. MicroRNAs are sequentially processed from longer precursor molecules that are encoded by the miRNA genes [1] (Fig. a Primary miRNAs (pri-miRNA) are transcribed from longer encoding DNA sequences (miRNA genes). The pri-miRNA contains one or more stem-loop structures of about 70 bases. including flowering plants.4 and let-7 were identified in worms. 1). fish. e. subsequently process the primary transcripts (or pri-miRNA) to generate mature miRNAs. This makes miRNAs one of the most abundant classes of regulatory genes in humans. frogs. mammals [38. 40. The encoding DNA sequence is much longer than the mature miRNA. more than 500 human miRNAs have been experimentally identified. 28]. MiRNA genes are referred to by the same name (termed mir) written in italics to distinguish them from the corresponding mature miRNA (termed miR) followed by a number. Subsequently. 71]. 75]. Fig. After untwisting. MicroRNAs are now perceived as a key layer of post-transcriptional control within the networks of gene regulation. The primary transcripts contain one or more stem-loop structures of about 70 bases. 41]. one RNA strand becomes the mature single-stranded miRNA. worms. the pre-miRNA is cleaved by the ribonuclease Dicer to generate a short RNA duplex [6. while the complementary strand. 1 The biogenesis and function of miRNAs. 1a). These small non-coding RNAs were named microRNAs (miRNAs) [38. 41]. and in humans [38. To date.g. Stem-loops are doublestranded RNA structures consisting of a nucleotide sequence that can fold back on itself to form a double helix with a region of imperfect base pairing that forms an open loop at the end (Fig. the fruit fly Drosophila. the ribonuclease enzyme Drosha . 68]. After export into the cytoplasm. Computational predictions of miRNA targets suggest that up to 30% of human protein coding genes may be regulated by miRNAs [46. and in single cellular algae and DNA viruses [66. MicroRNAs recognize their targets based on sequence complementarity [10].mir-1 or miR-1. is usually rapidly degraded (Fig. Drosha and Dicer. many more short regulatory RNAs were identified in almost all multicellular organisms.

1c).g. MicroRNA expression has been found to be deregulated in a wide range of human diseases including cancer. whereas messenger RNA levels may or may not be decreased. Their deregulation may therefore contribute to proliferative diseases such as cancer. it remains uncertain whether altered miRNA expression is a cause or consequence of pathological processes. Firstly. lin-4). To become effective. the mature miRNA forms a complex with proteins. Our group is currently investigating the role of miRNAs in mammary gland development and breast cancer pathogenesis. This base pairing subsequently causes inhibition of protein translation and/or degradation of the messenger RNA (Fig. miRNAs have been found to play a role in embryogenesis and stem cell maintenance [7]. cell proliferation. 42]. The potential mechanisms underlying this process were recently reviewed [30. miRNAs mainly inhibit protein translation of their target genes and only infrequently cause degradation or cleavage of the messenger RNA [1]. miRNAs have been suggested to contribute to the development of cancer [11]. A comparison of miRNA and gene expression identified miRNAs that classify molecular breast cancer subtypes [8]. Secondly. This review will focus on the connection between human miRNA biology and different aspects of carcinogenesis. hematopoietic cell differentiation [17]. In mammals. it was noticed that many miRNA genes were located at fragile sites in the genome or regions that are commonly amplified or deleted in human . the earliest miRNAs discovered in the roundworm C. 42].. when human miRNAs were discovered. In humans. growth control. neuronal differentiation. 67]. miRNAs regulate developmental timing (e. elegans and the fruit fly Drosophilawere shown to control cell proliferation and apoptosis [9. only very few miRNA target genes have been functionally validated. 60].complementary sites are usually within the 3′-untranslated region of the target messenger RNA. To date. Protein levels of the target gene are consequently reduced. Various techniques available to investigate miRNAs will also be discussed. The biological role and in vivo functions of most mammalian miRNAs are still poorly understood. As cancer is ultimately a consequence of disordered gene expression. and programmed cell death [9. Go to: MicroRNAs and cancer Three important observations early in the history of miRNAs suggested a potential role in human cancer. termed the RNA-induced silencing complex. 33. and brain development [59. knowledge of human miRNAs has been primarily descriptive. Although bioinformatics approaches can predict thousands of genes that are potentially targeted and regulated by miRNAs based on sequence complementarity. The underlying mechanisms of why and how miRNAs become deregulated are largely unknown. In invertebrates. The miRNA incorporated into the silencing complex can bind to the target messenger RNA by base pairing. However.

studied a well-known deletion on chromosome 13. the authors took advantage of a mouse model of human B-cell lymphoma. MicroRNAs as causal cancer genes at genomic breakpoints Five years ago. [27] demonstrated that additional expression of the mir-17-92 cluster accelerated c-Myc-induced tumorigenesis in mice. the miR-17-92 primary transcript was found to be overexpressed in tumor samples from lymphoma patients. were located within this 30-kb deletion. The Myc oncogene encodes the transcription factor c-Myc that regulates cell proliferation. mir-17-92. [13] found that two miRNA genes. This deletion had long been suspected to contribute to leukemogenesis. 24. They subsequently analyzed the expression of miR-15 and miR-16 in blood samples from patients with CLL. Further. The authors therefore suggested that mir-17-92 was the first potential non-coding oncogene. referred to as oncomir-1.cancer [14]. Thirdly. He et al. Both miRNAs were absent or downregulated in the majority (68%) of cases when compared to normal tissue or lymphocytes. [27] demonstrated that the miRNAs from the mir-17-92 cluster were overexpressed in lymphoma cell lines carrying this amplification. The mir-17-92 cluster—small RNAs with oncogenic potential A cluster of six miRNAs. the mir-17-92 cluster. These mice develop lymphomas due to an overexpression of the Myc oncogene. 63]. which is the most frequent chromosomal abnormality in chronic lymphocytic leukemia (CLL). The question remained whether the altered miRNA expression observed in cancer is a cause or consequence of malignant transformation. extensive studies had failed to identify a causal gene. and the Myc oncogenic pathway [27. and expression levels correlated with gene copy number of the mir-17-92 locus [27]. Calin et al. Two independent studies described the relationship between a miRNA cluster. mir-15 and mir-16. growth. He et al. and overexpression of c-Myc is common in cancer. However. In 2005. was found to be located within a region on chromosome 13 that is commonly amplified in human B-cell lymphomas [64]. malignant tumors and tumor cell lines were found to have widespread deregulated miRNA expression compared to normal tissues [12. and apoptosis. A third report demonstrated an interaction between let-7miRNA and the RAS proto-oncogene [32]. Calin et al. . 52]. three reports provided the first mechanistic insight into how miRNAs might contribute to carcinogenesis. This finding suggested that these two miRNAs were causally involved in the pathogenesis of chronic lymphocytic leukemia. To test their hypothesis that mir-17-92 actively contributes to lymphomagenesis. the first direct evidence for an involvement of miRNAs in cancer was reported [13].

thus. A single miRNA may regulate various unrelated target genes and thereby control opposing activities such as cellular proliferation and apoptosis. The same miRNAs may have oncogenic or tumor suppressor activity depending on the context and the cell type they are expressed in. mir-17-92. [32] showed that overexpression of RAS protein in lung cancer tissue correlated with reduced expression of let-7 miRNA. Nevertheless. [63] independently identified the same cluster of miRNAs. Mutations in the RAS oncogene are present in approximately 15–30% of all human cancers. . RAS proteins are membrane-associated signaling proteins that regulate cell growth and differentiation. Two reports demonstrated an anti-apoptotic effect of miR-17-92 through various pathways that promote cell proliferation and growth [55. They experimentally confirmed that let-7 can inhibit RAS expression in human cancer cell lines. The example of the mir-17-92 cluster highlights that a distinction between oncogenic and tumor suppressor miRNAs is likely to be an oversimplification. MicroRNAs with tumor suppressor potential The let-7 family of miRNAs was the first group of miRNAs shown to regulate expression of a proto-oncogene. to be regulated by the transcription factor c-Myc. Loss or reduction of let-7 in lung cancer leads to RAS overexpression. inhibit E2F1 expression. The authors therefore suggested a novel regulatory mechanism by which c-Myc fine-tunes gene expression by activating the transcription of target genes and by simultaneously inducing inhibitory miRNAs that reduce their translation. The mir-17-92 cluster which is also induced by c-Myc does. the pathology of the tumors indicated lower rates of apoptosis as compared to tumors with Myc overexpression alone. The authors therefore suggested that let-7 acts as tumor suppressor [32]. O’Donnell et al. the RAS protein. in contrast. A third study identified mir-17-92 as a mediator of angiogenesis in tumors induced by the oncogene c-Myc [19].The cellular function of miR-17-92 was not identified in these experiments. promoting cellular growth and contributing to tumorigenesis. Another group independently reported reduced expression of let-7 in lung cancers and found that this correlated with a poor prognosis [77]. and overexpression of the RAS oncogene is common in lung cancer. Johnson et al. A miRNA that controls expression of these potentially oncogenic proteins would be predicted to possess tumor suppressor activity. The ultimate function of a miRNA may depend on the tissue type they are expressed in and what target genes are present. The transcription factor Myc induces expression of E2F1 growth factor. 76]. Three recent studies contributed towards our understanding of the oncogenic potential of miR-17-92.

26]. also called “the guardian of the genome”. and in normal tissues [24]. such as let-7. The authors globally reduced the production of mature miRNAs through a knockdown of the miRNA-processing enzymes Drosha and Dicer in cell lines.Global loss of miRNA expression in cancer A global decrease in miRNA levels has been observed in human cancers. indeed. is the event that promotes malignant transformation. Global changes in miRNA expression may reflect the degree of cell differentiation [52]. Overall. Lu et al. The authors confirmed the finding that most miRNAs were expressed at lower levels in human tumorderived cell lines compared with the corresponding normal tissue [24]. MicroRNAs in the p53 tumor suppressor network Transcriptional networks are often deregulated in cancer cells and may lead to altered transcription of miRNA genes. To assess the effect of global miRNA loss in vivo. indicating that small RNAs may have an intrinsic function in tumor suppression. The p53 protein. these data clearly suggest that global miRNA loss enhances tumorigenesis. promote tumorigenesis. The Dicer mutant mice who had impaired miRNA processing developed an increased tumor burden. and normal tissues. Poorly differentiated tumors had lower miRNA levels compared with more-differentiated tumors. cancer cell lines. Cancers had significantly reduced global miRNA expression. miR-34. A recent study examined the expression of 241 human miRNAs in a comprehensive panel of human cancer cell lines. [37] proved for the first time that widespread reduction in miRNA expression does. Two recent studies identified a miRNA. They analyzed a total of 217 human and mouse miRNAs across 334 human cancers. considerable uncertainty remained as to whether the altered miRNA expression observed in cancer was a cause or consequence of malignant transformation. these cells generated faster growing and more invasive tumors compared to controls. These cells with global miRNA loss showed enhanced cellular growth in vitro [37]. as well as tumors which were less well differentiated compared to controls [37]. the NCI-60 panel. the authors deleted the miRNA-processing enzyme Dicer in a mouse model of lung cancer. with an expansion in tumor number and tumor size. Kumar et al. Earlier this year. regulates the cellular response to stress and cancer-initiating events such as . The mouse and human cancer cells consequently showed decreased steady-state miRNA levels. However. demonstrated that loss of miRNAs leads to upregulation of proto-oncogenes such as RAS and c-Myc. a study by Kumar et al. to be regulated by the p53 transcription factor [16. When injected into nude mice. The authors hypothesized that miRNAs can function to drive terminal differentiation and prevent cell division. it remains to be elucidated whether loss of all miRNAs is necessary or whether reduction of a subgroup of key tumor suppressor miRNAs. Until recently. [52] were the first to show that the expression levels of many miRNAs were significantly reduced in cancers compared to the corresponding normal tissues.

the overexpression of miR-17-92 correlates with amplification of its gene locus [27]. Similarly. A novel mechanism of miRNA regulation was suggested by Mayr et al. Regulation of miRNAs in cancer—who regulates the regulators? In few cases. mir-124a. high mobility group A2 (Hmga2). Together. the expression level of miR-10b in primary human breast carcinomas correlated with clinical progression [54].DNA damage. [26] found that a miRNA. decreased expression of miR-15 and miR-16 is associated with a corresponding chromosomal deletion [13]. MicroRNAs with a role in tumor invasion and metastasis Transcriptional networks may drive miRNA expression in cancers. As discussed above. In contrast. at least in some cases. Transcriptional or epigenetic regulation of miRNAs has been recently reported [53. Furthermore. Expression of miR-34 induces cell cycle arrest and thereby acts together with other effectors of the p53 tumor suppressor network to inhibit inappropriate cell proliferation. [56] and Lee and Dutta [44]. these data indicate that altered expression of miRNAs is not simply a secondary event that reflects the less differentiated state of cancer cells. suggesting that it is subject to epigenetic silencing through promoter hypermethylation. The transcription of a miRNA gene. the underlying cause of miRNA deregulation in cancer is clear. [54] suggested a model by which a pleiotropic transcription factor. suggest that specific miRNAs may have a role beyond the tumor-initiating event and directly participate in tumor progression and metastasis. Recent work from Ma et al. leading to tumor cell invasion and metastasis. Saito et al. induces expression of a specific miRNA. is directly activated by the transcription factor p53 after DNA damage. These findings. Both groups independently demonstrated that chromosomal translocations in a known oncogene. was shown to be inactivated by hypermethylation of its promoter in various human tumors. The expression of miR10b induced by the transcription factor Twist promoted cell migration and invasion in mouse and human breast cancer cells. miR-34. This process of epigenetic silencing is a wellknown mechanism to inactivate protein-coding genes in cancer cells and may similarly apply to miRNAs. They demonstrated that miRNA function could be regulated through loss of miRNA binding sites in the target gene. [73] demonstrated that miR-127 was highly induced in cultured human cancer cells after treatment with demethylating drugs. 73]. The miRNA genemir-127 is usually expressed in normal cells but not in cancer cells. He et al. if confirmed. Another group independently demonstrated that miR-34 is upregulated by p53 upon DNA damage and promotes apoptosis [16]. which suppresses its direct target and in turn activates a pro-metastatic gene. Twist. led to . miRNA expression is specifically driven by tumor suppressors and oncogenes.

Furthermore. there is evidence that miRNAs are regulated indirectly through control of their processing enzymes. Disrupted repression of Hmga2 by let-7 promoted oncogenic transformation and growth in mammalian cells. MicroRNA profiling—implications for cancer diagnosis Lu et al. their key advantage might be their high stability. a correct diagnosis could be established in 12 out of 17 of the tumors. These oligonucleotides need to be chemically modified to allow for stability in serum and cellular uptake. [81] showed that a downregulation of miRNAs in human cancer was not associated with reduced levels of the primary miRNA transcripts. Modified antisense oligonucleotides are already being developed to utilize the . Based on the differential expression of 217 miRNAs. they are long-lived in vivo [49] and very stable in vitro [78].000 messenger RNAs did not accurately classify the tumors [52]. In addition. A subgroup of gastrointestinal tumors.. tumors within a single cell lineage such as acute lymphoblastic leukemia were further differentiated according to their underlying genetic abnormality into BCR/ABL-positive tumors. e. [52] asked the question whether global miRNA expression profiles could classify human cancer. Thomson et al. gene expression profiling based on ∼16. 56]. and those with MLL gene rearrangement [52]. Cancers of epithelial and hematopoietic origin had distinct miRNA profiles.g. MicroRNA expression profiles clearly differentiated human cancers according to their developmental origin. Go to: MicroRNAs—novel therapeutic targets? Regulatory RNAs may also have therapeutic applications by which disease-causing miRNAs could be antagonized or functional miRNAs restored. Finally. T-cell tumors. the authors applied the miRNA expression profiles they had established to an independent series of 17 poorly differentiated tumors of unknown origin. These two studies provide the first evidence that disrupting the interaction of a single miRNA and its target can produce an abnormal phenotype in mammalian cells [44. In contrast. which might allow analysis of paraffin-embedded samples for routine diagnostic applications. This has potential important clinical implications. was distinguished by miRNA expression patterns. The most intuitive choice of molecules to correct altered miRNA–messenger RNA interactions are RNA oligonucleotides. The authors therefore suggested regulation of miRNAs during subsequent processing steps. In contrast to most messenger RNAs.loss of the let-7 miRNA binding sites in its messenger RNA. through altered function of the enzyme Drosha [81]. If miRNAs prove useful for clinical diagnosis. which arise from endoderm.

the “miRBase” database that provides up-to-date information on all published miRNAs [25]. This may represent a limitation for cancer therapies. then miRNA therapies may also be possible. Novel miRNA genes can be discovered by bioinformatics approaches searching for evolutionary conserved stemloop structures in the genome (reviewed in [3. Together. Any approach to knock down a particular miRNA with antisense oligonucleotides will only result in partial knockdown. If the delivery problem can be overcome. They developed miRNA inhibitors that can be transiently expressed in cultured mammalian cells. It remains to be seen whether indirectly mediated bystander effects on cancer cells that have not been directly targeted may partly overcome this limitation. The authors designed synthetic miRNAs to target overexpressed tumor proteins. these studies hold some promise of miRNAs as future therapeutic targets. a partial reduction of the disease-causing proteins in Alzheimer’s disease may lead to a clinical improvement and might be achievable by RNA based or miRNA gene therapy. A partial restoration of dopamine production by antisense therapy might result in a significant clinical improvement in Parkinson patients. A novel approach was recently reported by Ebert et al. Recent work by Krutzfeldt et al. 57]. such as Parkinson’s or Alzheimer’s disease. Both computational and experimental approaches indicate that many more miRNAs are likely to be identified [4. [36] demonstrated that modified cholesterol-conjugated antisense RNAs designated “antagomirs” could effectively inhibit miRNA function in vivo in the adult mouse. 5]). 51].intrinsic RNAi pathway for delivery of gene therapy. Similarly. Experimentally. Go to: Techniques and approaches to study miRNAs All known miRNAs are registered in a public web-based registry. miRNAs are discovered by cloning all small RNAs from a certain tissue type or developmental stage and subsequent sequencing to identify the subgroup of small RNAs that fulfill the criteria for miRNAs [38. A synthetic miRNA targeting HER-2 messenger RNA successfully inhibited HER-2 protein expression in ovarian cancer cells [83]. Two studies have successfully applied 2′-O-Methyl-modified antisense RNAs to inhibit miRNA function in cultured cells [29. [21]. The authors applied three daily intravenous injections of antagomirs and achieved effective inhibition of four miRNAs over a period of weeks in most tissues except brain [36]. One limitation of antisense RNA therapies is the restricted number of cells that can be targeted. a partial effect on function may be of therapeutic value in neurodegenerative diseases. such as HER-2 protein. A different approach was taken by Tsuda et al. [83]. which is reflected by the rapidly increasing number . In contrast. These competitive inhibitors termed “miRNA sponges” derepressed miRNA targets at least as strongly as chemically modified antisense oligonucleotides [21]. 5].

86]. conventional DNA microarray technology was modified to form miRNA microarrays.000 over the past 4 years [58]. In addition. Lu et al. in granulocytic differentiation [22]. 39.of annotated miRNAs which increased from less than 300 to more than 4. This approach allowed identification of a crucial role for a miRNA. MicroRNA expression studies Northern blot analysis is a well-established technique for studying messenger RNA expression and was soon adapted to detect miRNAs in cells or tissues [42. Functional characterization of miRNAs Various strategies have been used to investigate the function of specific miRNAs. In addition to mature miRNAs. denoting miRNA abundance [52]. 60. In worms and flies. these quantitative RT-PCR assays can be applied to analyze miRNA precursors and primary transcripts [31]. the modified antisense RNAs (antagomirs) described by Krutzfeldt et al. These allow for the analysis of miRNAs in small tissue samples or even single cells [79]. . loss-of-function mutants for specific miRNAs or miRNA families allow us to draw conclusions regarding possible physiological functions of miRNAs from the resulting abnormal phenotype [34. and labeling intensity. allowing for the detection of multiple miRNAs simultaneously across various samples [15. 61]. In situ hybridization for the detection of mature miRNAs has recently become possible by using special high-affinity locked nucleic acid (LNA)-modified DNA oligonucleotide probes and holds promise for the application on human formalin-fixed and paraffin embedded tissue [35. The solution hybrids are then analyzed using a multicolor flow cytometer measuring bead color. 82]. [52] developed a novel microarray strategy to improve probe specificity. The knockdown of miRNAs or pre-miRNAs using modified antisense oligonucleotides has proven particularly useful in cell lines [29. commercial assays for quantitative reverse transcriptase polymerase chain reaction (RT-PCR) have become available. denoting miRNA identity. LNA-modified antisense oligonucleotides have been successfully utilized to knock down specific miRNAs in cultured cells [22]. 84]. [36]. 57]. 62. They performed hybridization in solution using polystyrene capture beads that are coupled to oligonucleotide probes complementary to the miRNAs of interest. which is critical due to the short nature of mature miRNAs. which inhibit miRNA function in the adult mouse. as well as validation of microarray data. In parallel to microarray platforms. 45. Subsequently. may provide a potential research tool to study miRNA function in vivo. miR-223.

In mammals. In particular. 80].e. A common approach has been to express a miRNA in vivo while simultaneously expressing and monitoring the target messenger RNA linked to a reporter gene. induced defects in miRNA biogenesis are a useful tool for investigating the biological roles of miRNAs. This approach is based on the observation that some miRNAs can also downregulate messenger RNA levels in addition to downregulating protein levels of their target genes [49]. T helper cell differentiation and the germinal center reaction to produce a T-cell-dependent antibody response were defective [80]. exact base pairing between miRNAs and their targets commonly appears to be required only in the first six to eight bases from the 5′ end of the miRNA. 20].. Luciferase [10. MicroRNA target sites A validated biochemical strategy for identifying miRNA targets would be highly desirable. as loss-of-function mutants are not available for most miRNA genes. 10. and reported cardiac defects. An increasing number of sophisticated bioinformatics approaches are being developed to predict putative miRNA target genes [3. The fact that a single miRNA can regulate multiple targets . The short nature of this designated “seed region” allows a single miRNA to act on up to a hundred different target sites. 47]. and DGCR8 to study the consequences of a global decrease in mature miRNAs in cancer cell lines and in a mouse model for lung cancer [37]. 55. while mice lacking miR-1-2 had defects in cardiac morphogenesis and electrical conduction [88]. Two of the groups deleted the same DNA sequence for mir-155 and described severe immune defects [72. miR-1-2 and miR-208. A recent study utilized a combined knockdown of the miRNAprocessing enzymes Drosha. The two other groups deleted different miRNAs. deleted genes for single miRNAs in mice [72. Both approaches apply biochemical methods to purify the effector complexes of miRNAs associated with proteins and bound messenger RNA targets [2. 76]. Mice lacking miR-155 showed impaired function of B and T lymphocytes and dendritic cells [72]. Interestingly. A different approach to discovering miRNA target genes is to knock out or overexpress a particular miRNA and use conventional microarrays to identify genes that show changes in expression. Together. 85. Experimental validation of miRNA target sites has been limited to date. 18. 74]. four independent groups have. 80. Earlier this year. i. Two groups have recently reported promising approaches to experimentally identify miRNA targets. Dicer knockout mouse models have revealed essential roles for miRNAs in murine organogenesis [88]. 88]. 56. these two studies demonstrated a key role for miR155 in normal immune function. 69. Dicer1. for the first time. Mice lacking miR-208 showed inadequate cardiac growth in response to stress [85]. and all human miRNAs together may regulate up to one third of protein coding genes [10. This is based on the fact that miRNA target recognition is at least partly based on simple sequence complementarity.

.and a particular target may be regulated by various miRNAs suggests a highly complex network of miRNA-target interactions. Only 5 years after the first study reported a direct involvement of miRNAs in cancer. which is only beginning to be unraveled. these small RNAs have already significantly improved our understanding of carcinogenesis. Go to: Conclusions Over recent years. we will have to take into account miRNAs and their regulatory networks if we aim to understand the complex processes underlying malignant transformation. miRNAs have emerged as major players in the complex networks of gene regulation and have been implicated in various aspects of human disease. In addition to proteincoding oncogenes and tumor suppressor genes.