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INTRODUCTION

1.1.Cancer
Cancer is a class of diseases or disorders characterized by uncontrolled division of cells and the ability of these cells to spread, either by direct growth into adjacent tissue through invasion, or by implantation into distant sites by metastasis (where cancer cells are transported through the bloodstream or lymphatic system). Cancer arises from cells in the body which were once normal cells. In the growth of normal cells, a finely controlled balance exists between growth- promoting and growth-restraining signals such that proliferation takes place only when required. This order is tilted only when more cells are required such as in wound healing. In this situation differentiation takes place in an orderly manner and proliferation ceases when no longer required. However, in tumour cells this process is disrupted, cell proliferation happens continuously and loss of differentiation can occur. The normal process of programmed cell death may cease to operate These cells which are now “transformed” to grow and divide and keep dividing in an uncontrolled manner. They differ very subtly at first compared to the cells of the normal tissue from which they originate

A tumor or tumour is the name for a neoplasm or a solid lesion formed by an abnormal growth of cells (termed neoplastic) which looks like a swelling. A tumor can be benign, pre-malignant or malignant, whereas cancer is by definition malignant Two types of tumors exist, benign and malignant. Benign tumors are not cancerous. They can usually be removed by surgery and generally don't grow back The cells in benign tumors don't spread and it is rare for a benign tumor to be life threateningMalignant tumors, on the other hand, are cancerous. The cells in them are abnormal and divide randomly and chaotically. The cells behave aggressively and attack the tissue around them.

They also can jump away from the malignant tumor and enter the bloodstream or lymphatic system to form new tumors in other parts of the body. This type of spread is known as metastasis.

1.2. Origins of Cancer
All cancers begin in cells, the body's basic unit of life. To understand cancer, it's helpful to know what happens when normal cells become cancer cells. The body is made up of many types of cells. These cells grow and divide in a controlled way to produce more cells as they are needed to keep the body healthy. When cells become old or damaged, they die and are replaced with new cells. However, sometimes this orderly process goes wrong. The genetic material (DNA) of a cell can become damaged or changed, producing mutations that affect normal cell growth and division. When this happens, cells do not die when they should and new cells form when the body does not need them. The extra cells may form a mass of tissue called a tumor

Fig 1:Difference between Normal and Cancer Cell Growth

* Benign tumors aren't cancerous. They can often be removed, and, in most cases, they do not come back. Cells in benign tumors do not spread to other parts of the body. * Malignant tumors are cancerous. Cells in these tumors can invade nearby tissues and spread to other parts of the body. The spread of cancer from one part of the body to another is called metastasis.

Fig 2:Benign and malignant tumors

.1.3.

Classification:

Cancers are classified by the type of cell that resembles the tumor and, therefore, the tissue presumed to be the origin of the tumor. These are the histology and the location, respectively. Examples of general categories include:

Carcinoma: Malignant tumors derived from epithelial cells. This group represents the most common cancers, including the common forms of breast, prostate, lung and colon cancer.

alteed nuclear. Many of these tumors are most common in children. and young children most often found on the body midline. 1. • • • Blastic tumor or blastoma: A tumor (usually malignant) which resembles an immature or embryonic tissue. abundant mitosis ● Often grow over one another and form multilayers. In adults most often found in the testicle and ovary. . particularly at the tip of the tailbone.BIOCHEMICAL CHANGES ● Increased synthesis of DNA and RNA ● Show increased rate of glycolysis ● Show alterations of permeablity and memberane charge ● Changes in composition of glycoproteins and glycosphingolipids on cell surfaces ● Apperarance of new antigens and loss of certain antigens.MORPHOLOGY OF CANCER CELLS ● Larger than normal cells ● Show nuclear and cellular polymorphism ● Hyperchromatism. or mesenchymal cells.4. babies.5. cytoplasmic ratio.• Sarcoma: Malignant tumors derived from connective tissue. ● Diminished adhesion 1. Lymphoma and leukemia: Malignancies derived from hematopoietic (bloodforming) cells Germ cell tumor: Tumors derived from totipotent cells. in horses most often found at the poll (base of the skull). in fetuses.

promotion. chemical and viral agents that lack the capacity for initiation and promotion may actively convert the cells in the stage of promotion to the stage of progression Fig 3:Mechanisms of carcinogenesis . radiation. Carcinogenesis can be actively caused by chemicals. The final stage of carcinogenesis. which interferes with biological processes.6.. i. progression. The mechanism of the induction of carcinogenesis may be due to alteration in the genomic structure. and infectious biological agents.1.Carcinogen Carcinogens are agents that causes cancer or increase the risk of getting cancer by altering cellular metabolism or damaging DNA directly in cells.e. and induces the uncontrolled. malignant division. In addition. ultimately leading to the formation of cancer tumors. The action of specific carcinogenic agents depends upon the stages of cancer development such as initiation. and progression. may occur spontaneously enhanced by the formation and propagation of genetic errors.

1. Proto-oncogenes can be mutated by carcinogenic agents to become oncogenes. Cancers result from a series (progression) of gene mutations that typically involve two categories of function: promotion of cell division and inactivation of cell cycle suppression.7.Molecular Mechanisms of Cancer: Fig 4:Various Stages of Cell Cycle Cancer is a disease that begins in the cells of the body. the cells grow and divide as the body needs them. no less. No more. Proto-oncogenes are normal genes that promote cell growth and mitosis. In normal situations. whereas tumor suppressor genes discourage cell growth. These extra cells lump together to form a growth called tumor. This orderly process is disturbed when new cells form that the body was not needed and old cells don't die when they should. Oncogenes produce excessive levels of growth promoting proteins .

such as thrombosis or hormonal changes. immortalization due to over expression of telomerase. hemorrhage (bleeding). whereas suppressor genes are recessive. They contain loss-of function mutations. loss of contact inhibition. Compression of surrounding tissues may cause symptoms such as jaundice).Tumor suppressor gene products typified by p53 are frequently transcription factors that suppress mitosis and cell growth to allow for DNA repair. Only one copy of a proto-oncogene needs to mutate for gain-of-function. Nearly half of all cancers involve altered p53 genes. specific conditions that are due to an active cancer. Although advanced cancer may cause pain. fracture of affected bones and neurological symptoms. fatigue and cachexia excessive sweating anemia and specific paraneoplastic phenomena. poor appetite. . increased cell division. it is often not the first symptom. p16/CDKN2A and BRCA (breast cancer susceptibility protein) types 1 and 2. 1. Both copies of a suppressor gene need to mutate to cause loss-of-suppressor function. bone pain. Cancer results from cumulative mutations of proto-oncogenes and suppressor genes which together allow the unregulated growth of cells. Over time malignant cells can self-select for characteristics that make them more malignant: ability to avoid apoptosis. pain and ulceration. become metastatic.e. TP53. • Systemic symptoms: weight loss.8. hepatomegaly. and able to promote angiogenesis. • Symptoms of metastasis (spreading): enlarged lymph nodes. SMAD4. APC (adenomatous polyposis coli). growth-factor self-sufficiency and resistance to anti-growth factors. Oncogenes are typically dominant because they provide gain-of-function. i. Other suppressor genes include Rb (retinoblastoma family). • Local symptoms: unusual lumps or swelling (tumor). cough and hemoptysis. Mutations of tumor suppressor genes can be inherited. altered differentiation.Symptoms of cancer: Symptoms of cancer metastasis depend on the location of the tumor.

Radiation therapy can be administered externally via external beam radiotherapy (EBRT) or internally via Brach therapy.Existing lines of treatment: Conventional cancer treatment can include surgery. e.g. . Chemotherapy: Chemotherapy is the treatment of cancer with drugs (anticancer drugs) that can destroy cancer cells. Basically. and biological therapy . These cells usually repair themselves after chemotherapy.9. When the cancer has metastasized to other sides in the body prior to surgery. Docetaxel. chemotherapy agents kill cancer cells that are actively multiplying The general disadvantage of chemotherapy. Most forms of chemotherapy target all rapidly dividing cells and are not specific for cancer cells hence the chemotherapy has the potential to harm healthy tissue especially those tissues that have a high replacement rate. but this is not always possible. with the duplication of the DNA or the separation of newly formed chromosomes. Estramustine or others. chemotherapy. Radiation therapy: Radiation therapy is the use of ionizng radiation to kill cancer cells and shrink tumors. Vinorelbine. Surgery: Cancer can be cured if entirely removed by surgery. It interferes with cell division in various possible ways. complete surgical excision is usually impossible. mono clonel antibody therapy. The effects of radiation therapy are localized and confined to the region being treated.. Chemotherapy is a treatment based on action of several agents such as: Mitoxantrone. is the drugs cannot discriminate between fast-growing cells and kills all cells whether they are part of controlled or uncontrolled process. no matter of type of cancer.1. hormone therapy. radiation.

At the same time.10. Foscan. it has been proposed that a drug which inactivates telomerase might be effective against a broad spectrum of malignancies. and would function normally in its absence.Plant source as tumor remedy Plant products have been used for centuries as medicines. with recipes handed down from generation. Hormonal suppression: The growth of some cancers can be inhibited by providing or blocking certain hormones. Tookad. most healthy tissues in the body express little if any telomerase. WST09. Common examples of hormone-sensitive tumors certain types of breast and prostate cancers. Photofrin and Visudyne) which are triggered by light of a specific wavelength. also known as PDT. They are available and . Vaccines to generate non-specific immune responses are subject of intensive research for a number of tumors notably malignant melanoma and renal cell carcinoma. Today in most prevalent treatments. and use of interferon and interleukin. uses photosensitive drugs (such as 5-ALA. Photodynamic Therapy. Photodynamic Therapy: Photodynamic Therapy is generally a non-invasive treatment using a combination of light and a photosensitive drug.Immunotherapy: More contemporary methods for generating non specific immune response against tumor include intraversial BCG immunotherapy for superficial bladder cancer. Metvix. WST11. 1. Telomerase therapy: Most malignant cells rely on the activity of the protein telomerase for their immortality.

Proper clinical trials were also not conduced. which may contain phytochemicals that can modulate gene expression inhibit carcinogenesis via multiple pathways. Many are secondary metabolites. and herbivores. Each herb may hold active chemicals.are less costly then allopathic medicine. these substances (particular the alkaloids) serve as plant defense mechanisms against predation by microorganisms. Many of the herbs and spices used by humans as season food also yield useful therapeutic compounds with only a few exceptions. This is known as “Ganakasika” botanically equated as Premna integrifolia Linn. Many plants synthesize substances that are useful for the maintenance of health in humans and other animals. . insects. and there is generally a more culturally sensitive attitude on the part of these practitioners. most herbal treatment have not been tested for safety and efficacy utilizing scientific parameters. Many studies revealed reduced incidences of many common forms of cancer if treated with herbs. These include aromatic substances. This plant source is used by Ayurvedic practitioners for various preparation that controls several oilments. most of which are phenols or their oxygen-substituted derivatives such as Tannin. Ethanolic extract of the aerial portion of this traditional drug source is scientifically validated for its anticancer potential. practitioners are available.000 have been isolated – a number that is estimated is to be less than 10% of the total. of which at least 12. In the present dissertation a commonly available plant belonging to Verbinaceae is selected and screened for the its anticancer potential. In many cases.

REVIEW OF LITERATURE Two novel tritrepene acids. 30-dihydroxy-lup-20(29)ene. (4)3beta. (1)beta-sitosterol.03 (μg/ml) against P-388 cell lines. ZK etal.. Frculontonic acid A(1) and B(2) as well as the extract of leaves.butulin.. together with known flavone. These compounds together with eleven terpenoids previously isoalated from Euphorbia lagascae and E. 2009) (3)dibutyl phthalate.The structures of two new compounds were established on the basis of 1D and 2D NMR spectroscopic data interpretation and chemicals conversions.tuckeyana were tested for their apoptosis induction activities by flow cytometry on L6178 human MDR1 gene-transfected lymphoma and result showed moderate apoptosis induction effects. The compound 5 should potent anticancer effects against KB cells (Dai .. Among them confusameline was found to be most cytotoxic isolate and exhibited most potent activity (ED50=0.HA-29 and A 549 cell lines in vitro . (5)daucosterol..(Duarete N etal. 2004) Seven known pentacyclic triterpenes and one steroid were isolated from Euphorbia lagascae methanolic extract and identified by physical and spectroscopic methods. 2004) Melicarpine. stems and twigs of Manihot esculenta studied for cytotoxic activity . 2009) The petroleum ether fraction of Buxus microphyla was subjected column chromatography. (2)stigmasterol. (Chaturvedula VSP etal.ayanin were isolated from the leaves of Melicope somecarpifolia All the compounds along with 15 compounds isolated previously from the same plants were tested against P-385. (Chenm JJ etal. The two new compounds showed moderate cytoxicity against the A2780 human ovarian cancer cell lines. Eight components were identified namely. Semecarpine and (+)-8-methoxyplanty desmine.

2009) The study aimed to at evaluate the cytotoxic mechanism of Calotropin on human chronic myeloid leukemia K562 cells.prostate and breast cancers. a natural components of ginger exhibits antiinflamatory and anti tumorigenic activities. The leukotriene A(4) hydrolase (LIA(4)H) protein regarded as a relevant target for cancer therapy.. Respectively leukemia. Gingerol suppresse anchorage independent cancer cell growth by inhibiting LTA(4)H activity in HCT 116colorectal cancer cells.A flavanone has been isolated from Bauhinia variegate and its structure was identified by colour reactions and spectral analysis.. ovarian.2009) Gingerol. Despite its potential efficacy in cancer.renal. colon.central nervous system maianoma..(Wang SC etal. Selectively in androgen receptor (AR) positive prostate cancer cells and shifted the propotion in each phase of cell cycle towards(2)-M phase in AR negative prostate cancer cells. 2009) . Exposure of prostate cancer cells to W. In silico prediction using a reverse –doking approach revealed that LTA(4) H might be a potential target of gingerol. It up regulated the expression of p27 leading to this arrest by down regulating theG2/M regulatory proteins cyclins A and B. The isolated flavanone was tested for cytotoxic activity against 57 human tumour lines.and by up regulating the cdk inhibitor.chinensis extract induced apoptosis pathway. the mechanisms by which -gingerol exerts its chemopeventive effects remains unclear. The results showed that the flavanone showed cytotoxic activity against human tumour cell lines. non-small cell lung. Calotropin inhibited the growth of K562 cells in a time and dose-dependent manner by G(2)/M phae arrest.. (Tsai CH etal.it down regulated anti-apoptotic signalling (XTAP and surviving) and survival pathways (P-Akt and NFKappaB) leading to caspase-3 activation which resulted in the induction of apoptosis. (Jeong CH etal.(Rajkapoor B etal.p27furthermore. 2009) The Invivo efficacy and mechanisms of action of oral administrations of a standardized extract of W.chinensis were analysed in animals bearing a subcutaneous orthiopic prostate cancer xenograft.

(Huang GC etal.4-trihytrochlcone. 2009)... Antihelmintic and anticancer activities were evaluated by the inhibiting concentration at 50% death.(IC(50) and the selectivity index(ST) relative to human fibrbiasis. 2005) Casuarinin a hydroluzable tannin isolated from the bark of Terminalia arjuna was investigated for its antiproliferative activity in human breast adenocarinoma MCF-7 cells. ..(Atjanasuppat K etal.50µg/ml..(kuo PL etal.Human amelanotic metanoma) and humancervical carcinoma (Hela)by the SRB assay. Cytotoxicity of the extracts was evaluated against two cancer cell lines.2009) The methanolic extract of leaves of Cassia tora (CTME) showed potent lipid peroxidaxlon inhibitory activity as well as showed potent antiproliferative and apoptosis inducing activity against HeLa cell lines. Strong cytotoxicity was shown by the methylene chloride extract of Michelia champaca bark and the methanol extract of Carcama longarhizome against C32 and Hela cell lines respectively.2.(Yang ZG etal. with an IC50 of 10. 2005) Evodiamine isolated from Evodia rutaecarpa showed Antiproliferation and apotosis inducing activity against human colorectal carcinoma cells(colo-205). The results showed DNA fragmentation in p-38801 bearing CDF1 mice at a dosage of 5mg/kg body weight . Isoliquiritigenin (4. The result showed that casuarinin indues the apoptosis in MCF-7 cells..( Rejiya CS etal.ISL)is a natural pigment with a simple chalcone structures.study discovered on antitumour bioactive compounds from That indigenous plants.(Hso yl etal. The cell growth inhibition achieved by ISL treatment resulted in programmed cell death manner.. 2005) The anti-tumour activity of the diethyl extract of zingiber zerumber was measured in cultured P-388DI cells and on animal model of P03881bearing CDF mice. 2009).

a triterpene saponin isolated from Gleditsia sinensis. Administration (i. E.25 lymphoma and Melanoma.B. 125 extracts in total.5 µg/ml in dose and time –dependent manners.patullum was tested against HEp-2 cell lines the results that the both plant extraction showed moderate cytotoxic effect against HeLa cell lines. 2003) . showed significant cytotoxicity against BEL-7402. (Syu CM etal.. HELA.five species. BGC-823.mysorense and H.ochracea. The methonalic extracts in the H. lappadilactone.”Induction of apoptosis and G2/m cell cycle arrestare the mode of action of Gledisioside E .5178-m. 2004) Aqueous and organic extracts of Asteraceae (compositae) collected from the state of Rio Grande do sul. OVGPR-3 and HeLa cell lines.p) of 2 or 10 mg/kg of magnolia significantly suppressed liver and spleen metastasis. These data suggest that magnolia possess strong anti metastatic ability and that it may be a lead compound for drug development. (Monks NR etal. 2004) Gleditsioside E. Eupatorium macrocephalum.small cell lung cancer cells. Twenty.pedunculosum and Stenachaenium niedeli all produced IC 50 values below 5 mg / dl. Bioassay –direced fractionation of Saussurea lappa lead to the isolation of a novel lappadilactone and some sequiterpene lactones as cytotoxic principle against selected human cancer cell lines. were screened against HT29 human colonadenocarcinoma cells and Ncl-H460 human non... dehydro-costuslactone and cortunolide exhibited the most potent cytotoxicity with CD50 values in the range 1:6-3.HL-60 and MCF-7 cell lines. Brazil were tested in vitro or cytotoxic activity against human solid tumour cell lines.Vijayan etal 2004 reported the in vitro cytotoxicity and anti-tumour properties of Hypericum mysorense and Hypericum patulum. (Ikeda k etal.. The cytotoxicities were not specific and showed similar activities against HepG2. 2009) The anti metastatic effects of Magonolia isolated from Magnolia oborata were evaluated by an experimental liver and spleen metastasis model using 1.(Zhong L etal. Extracts Bacchoris cordifolia.

is screened for its antitumour potential. Poochendura pattai a traditional drug which is botanically equivalted as Plectranthus urticifolius Hook. 2004)..3dimethoxy flavone. ladanein and hispidulin were isolated from the methenolic extract of the Cerial parts of Artemisia argyi and structure compounds were elucidated on the basis of their structural data. compound 2 inhibited proliferation of a couple of tumor cell lines ( Leo JM etal.5.0mg/ml .. Packed cell . Among them alpha mangostion showed complete inhibition at 10µ/m through the induction of apotosis.the extract induces the DNA transformation . 5.2000) Embelin a naturally occurring benzoquinone isolated from Embelia ribes showed significant antitumor activity by increasing the life span and reversing the elevated levels glycorportein in tumor bearing rats.5trimetoxy flavone. .(Matsumoto K etal. 2005) The effect of six xanthones from Garcinia mangstana were examined on the cell growth inhibition of human leukemia cell line HL60 All xanthones displayed growth – inhibitory effects.6-dihydroxy-7.The methonalic extract of pereshipa bleo showed cytotoxic activity against T-47D cell with Ec 50 of 2. 2003) The 7.7-tetramethoxy flavones..6..3-trihydroxy-6.7. and gave encouraging results (Brindha P etal..3-4-trimethoxy flavones.8 –dihytroxy flavanone isolated from the seeds of Alpinia katsumadai was found to have an in vitro cytotoxic effect against A 549 (a human cancer cell lines) and k562 (a human leukemia cell line. 5-hydroxy-3-4 6. F.. 2004) In EAC tumor induced mice the methanol extract of Mucuna pruriens at the doses of 125mg and 250mg/Kg.4-trihydroxy-7. These flavanones inhibited farnesyl protein transferees with IC50 values of 25-200 µg/ml.chormarin Marigination which results in the activation of apotosis in T-47D cells(Tan ML etal. (Hahm ER etal. (Chitra M etal. 2005) The flavanones 5.4. bw significantly inhibited the tumor volume.

2005) Ursolic acid pentacyclin triterpene component was screened for anti cancer potential against endometrial cancer cell line SNG – II. Ehrlich Ascites carcinoma. 1974) Administration of Black Tea to EAC bearing Swiss albino mice caused significant decrease in the tumor cell count and increased life span in dose dependent manner. ( Chakrabortui etal. It was found that the ursolic acid strongly inhibited the SNG II cell lines in dose and time dependent manner. Sarcoma E 0771. and mamaray adeno carcinoma cell lines in animal models..volume and cell count and brought back the hematological parameters ( Rajeswar etal. Belonging to the family Verbenaceae was selected and the ethanolic extract of the plant material was screened for anticancer activity against Ehrlich Ascites carcinoma cell lines. Hence in the present study a common plant “Munnai” botanically equated as Premna Integrifolia Linn. . 2003) From the literature we found that the plant drugs plays a significant role in the development of anticancer drugs. ( Arindan Bhattacharyya etal... Mechanism behind this was found by activity of apoptotic pathway which decrease the BCL-2 protein 20(anti apoptotic factor) and increased BAX protein expression ( Apototic factor) (Yumiko Achiwa etal. 2005) Withaferin A isolated from Withania somnifera showed marked tumor inhibitory activity in carcinoma Nasopharynx. Sarcoma 180..

Many treatment modalities are available for cancer at present but they are always associated with side effects. belonging to the family Verbenaceae was selected and the ethanolic extract of the plant was screened for anticancer potentials against Ehrlich Ascites Carcinoma cell lines.AIM AND OBJECTIVES Cancer is one of the deadliest as well as induced disease characterized by uncontrolled and unwanted growth of cells leading to the dysfunction of the organs where it proliferates. Hence need of the hour is to develop an anticancer drug which is cost effective. Studies were also carried out from standardization point of you. To determine the Chemical Standards  To carry out phytochemical screeing for various extracts. The main objectives of the present study are:  for the selected plant drug. human compatible with proven anticancer effect. Based on the literature review.  To conduct In-Vitro Cytotoxic studies against Ehrlich Ascites Carcinoma cell lines. present investigation aims at to identify and develop a novel anticancer herbal drug.  To conduct in-depth out In-Vivo anticancer studies against Ehrlich Ascites Carcinoma cell lines . Premna integrifolia Linn. Traditional plants might provide useful sources for developing new anticancer drugs and could be a good alternate to existing lines of cancer therapies.

17 30 20.75 Viable cells Viable cells (%) 95.68 20 10 0 Control 50 100 μg/ml 4.94 4 30 37 52 93 Death cells Death cells (%) 4.6 40 32.4 45.68 32.32 76.06 Dead cell = Accepts trypan blue dye Viable cell = Not accepting trypan blue dye Fig 6:SHORT TIME INVITRO CYTOTOCITY STUDY 60 53.79 79.21 250 500 .21 20.Table 7-Cytotoxic Effect Of EEPI on EAC cells (Trypan Blue method) Concentration of EEPI (μg/ml) Control 50 100 250 500 91 95 78 76 79.60 53.17 40.83 59.06 50 percentage of cell death 40.

06% of cell death. .(EEPI) against EAC cell lines was Tabulated and graphically represented in (Table 7 & Fig 6). The percentage of cytotoxicity was found to increase with increase in the concentration of EEPI. .In –vitro cytotoxic effect of the Ethanolic extract of Premna integrifolia Linn. At minimum concentration (50 µg/ml) showed 20.68% cell death where as the higher concentration (500 µg/ml) showed 53.

) EAC control + EEPI (200mg/kgbw.Table 8-Effect of EEPI on survival time of the Tumor bearing mice Particulars EAC control EAC control + EEPI (100mg/kgbw.15* Fig 7: Effect of EEPI on survival time of the Tumor bearing mice .1 24±1.01statistically significant when compared to EAC control group EEPI: Ethanolic Extract of Premna integrifolia EAC: Ehrlich Ascites Carcinoma Mean survival time (MST) (days) 19±1.63* 63.31 52.59* Increase in life span(%) (ELS) 26.) EAC control + 5FU (20mg/kg bw) Values are Mean ± S.71* 31±1..E.37 29±2.M. n=6 *p<0.

15% Table 9 –Effect of EEPI treatment on tumor growth . On treatment with various concentration of EEPI. The animals which received the 200mg/kg body weight of EEPI showed 52.31 52.63% of increase in the life span of tumor bearing animals (Fig 7). The standard drug 5-flurouracil increased life span up to 63.63 63.15 Group V Table8 indicates the increase in the life span of tumor bearing animals.Increasing in life span% 70 60 50 40 30 20 10 0 0 Group II GroupIII Group IV 26.

D.5±0.4±0.36* Viable cell 10 10 cells/L 4.40 4.10 1.49 2.41 0.2±0.32±0.7±0.21* 2. n = 6 *p<0.13* 1.34* Non-viable cells 1010 cells/L 0.7±0.27* 0.7±0.21* Group I Group II Group III Group IV Group V Values are Mean ± S.07 0.9±0.32 1.1±0.particulars Tumor volume(ml) 1.9±0.05statistically significant when compared to Group II Group 1: Normal control .1±0.5±0.

Fig 8: Effect of EEPI treatment on tumor growth 2 1.6 0. of EEPI. Group 4: EAC + 200 mg/kg bw.5 1.1 1.4 0.Group 2: EAC control Group 3: EAC + 100 mg/kg bw.5 .4 1.8 0.8 1. of 5-fluoro uracil.7 0.2 1 0. Group 5: EAC + 20 mg/kg bw.6 Tumour volume (ml) 1.2 0 Group I Group II Group III Group IV Group V 0. of EEPI.

2±0. n = 6 *p<0.8±0.48* 15.D.7±0.2* 103 cells/mm3 6. of EEPI.The EAC inoculated mice showed increased Ascites fluid volume (1.3*6.3 7.3±1.1±0.3±0.5±0.4±0.32) and increased viable cell counts (Table 9).9±0.27) (Table )decreased viable cells count and increased non.3±0.4 13.4 11.3±. .1±0.7* WBC Values are Mean ± S.0.05statistically significant when compared to Group II Group 1: Normal control Group 2: EAC control Group 3: EAC + 100 mg/kg bw.3±0.5 4.5 8.3* 5. On oral treatment with two different concentration of EEPI administration for 14 days decreased the Ascites fluid volume group III (1.41) and group IV (0.3±0.3 4.1±1.35* 106 cells/mm3 5.viable cells counts of tumor bearing animals (Fig 8) TABLE 10 -Effect of EEPI on Hematological parameters RBC count Hb Particulars Group I Group II Group III Group IV Group V (g %) 15.57 14.8 9.7±0.6±0.3 5.7±0.

Death rate due to cancer is keeps on increasing. Need of the hour is to find a safe.Group 4: EAC + 200 mg/kg bw. However most of the modern therapies available for cancer treatment at present are capable of producing serious side effects.Not more then 3% . Group 5: EAC + 20 mg/kg bw. and more than 11 million people were diagnosed as cancer patient every year. in the present study physico and phyto chemical standards were determined for the plant drug selected for the study. of 5-fluoro uracil.Not more then 2% . cost effective and efficacious drug for this deadly disease. PURITY AND STRENGTH: (1) Foreign matter(%) (2) Total ash content (%) (3) Water soluble ash (%) . employing both In-Vitro and In-Vivo methods.Not more then 4% . of EEPI. As standardization plays a vital role in herbal drug development. belonging to the family Verbenaceae was selected and screened anticancer potential against Ehrlich Ascites Carcinoma cell lines. DISCUSSION Cancer is one of the second largest killer disease in the world. TEST FOR IDENTITY. In the present study with a view to develop a safe a plant anti cancer herbal drug botanically equated as Premna integrifolia Linn.

and Steroids in the plant drug powder Under study. The phenolic content may be attributed to the therapeutical potential of the plant drug under study. Lignin and Quinones. Coumarins. (EEPI) was tested against Ehrlich Ascites Carcinoma (EAC) cell lines using Trypan Blue dye exclusion method. Phytochemical screening of drug powders as well as various extracts confurmed the presence of Alkaloids. This indicates the presence of Alkaloids. Steroids. The cytotoxic potential of Ethanolic Extract of Premna integrifolia Linn. The organic constituents such as carbohydrates. At the concentration 500 µ g /ml EEPI showed .Not less then 2% .Not less then 11% - Not less then 9% Not less then 18% Fluorescence analysis of the plant drug revealed the presence of Brown.Not more then 1% . protein and phenol were found to be higher in the plant material.(4) Acid insoluble ash (%) SUCCESSIVE EXTRACTIVE VALUES: (1) Hexane (%) (2) Chloroform(%) (3) Ethyl acetate(%) SOLUBILITY: (1) Alcohol (%) (2) Water(%) . The preliminary experiment was carried out using five different concentration of test drug.Not less then 2% . Flavanones.and UV light showed various shades of green fluorescence (Table3 ). Yellow and Green fluorescence in Day light. Flavones.

.1958). Administration of two dose levels of EEPI to the EAC animals increased the blood glucose level. The present study EEPI treatment reduced the WBC count. Hence the test drug might be considered as a good anticancer drug.63% (Table 8). The increased WBC count of tumor bearing mice are due to immune response against cancer cells. The ascites fluid is essential for tumor growth because it constitutes the essential nutritional source for tumor (Feccchi OD etal. The ascites fluid volume will directly represent the tumor growth. The criteria for good anticancer drug is to reduce the WBC count. . Hence the test drug may consider as good anticancer drug.06% of cell death whereas low concentration as much as 50 µ g/ml showed 20. (Price VE etal.This may be due to the excess hemolysis. cells were permeable to the Trypan Blue. The reliable criteria for judging the value of any anticancer drug is prolongation of life span of tumor bearing animals. In the presence study EEPI effectively reduced the ascites fluid volume this clearly indicate that the test drug provides the direct cytotoxic effect to the tumor cells or may inhibit the vascular permeability and cellular migration. (Table 7) As the part of the apoptosis precede the loss of membrane integrity. The decreased serum protein content of EAC animals is due to the functional impairment of hepatic tissue. (Clarkson BO etal. This indicates that the EEPI are capable of protecting the hemopoietic system.53.. On treatment protein levels found to be increased. In cancer therapy major problem encounted are (RBC) myelosuppression and anemia.1990).1965) In the present study EEPI increased the life span of tumor bearing animals up to 52. The lowered blood glucose hypoglycemia was reported in the EAC animals by (Baliant Z 1191).. This indicated that the plant drug suppressed the solid tumors by blocking the energy fuel of cancer cells. On treatment the RBC and hemoglobin levels were reverted back to normalcy. This indicated that the plant drug protect the hepatic tissue.68% of cell death.

SUMMARY AND CONCLUSION Cancer is one of the deadliest genetic as well as induced diseases resulting due to the defects in the genetic material. 1971). Chemotherapy. But these lines of treatments are known for their serious . and Monoclonal antibody therapy.glycoprotein component and antioxident system.reduced tumor growth and restoration of the biochemical . This is because of the loss of cell growth control mechanism caused by various carcinogenic agents. This may be due to increased lipolytic activity. On treatment with test drug the increased level of DNA & RNA were restored to normal level. EAC inoculated mice showed increased amount of nucleic acid content such as DNA & RNA (Gkazer RT etal. on treatment with EEPI the serum cholesterol and Triglycerides were found to be decreased. This may be due to the increased cellular proliferation and . In the present study oral treatment of two different concentrations of EEPI (100&200 mg/kg bw) to the EAC induced animals resulted in increased life span .The induction of cancer resulted in the increased serum Cholesterol and Triglycerides levels. This indicates that the test drug may block the lipolysis and increased the lipogenesis..The data of the results obtained in the present study were comparable with the result obtain on treatment with the standard drug 5-FU and it was found that the dose level (200mg/kg bw) of EEPI was found to be the effective dose. Today modern medicine provides number of conventional methods such as Radiation therapy.hepatic marker enzymes . It is characterized by uncontrolled and unwanted growth of cells. This might be due to blocking of proliferation of tumor cells at G1 phase of cell cycle and inhibiting the activity of polymerase enzymes. Hormonal suppression.

Based on the literature review. in the present dissertation a common traditional plant source Premna integrifolia Linn.side effects. belonging to be verbenaceae family was selected and ethanolic extract was screened for anticancer activity against Ehrlich Ascites Carcinoma cell lines. Plants and plants derived products have been widely accepted as a good therapeutic agents with lower side effects particularly in the management of cancer. .

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