CANCER GENETICS 3

Alternative Pathways to Cancer

The paths that cells take on their way to becoming malignant are highly variable. Within a given cancer type mutation of particular target genes such as ras or p53 may be found in only a subset of otherwise histologically identical tumors.

In familial cancer cases, the cancer occurs more frequently in the family than in the population they already have. Onset is also frequent as already 1 copy is inherited, 1 more has to be acquired. In Familial Adenomatous Polyposis when 2 copies are mutated then benign tumor becomes malignant. Familial cancer suggests a clustering of cancers that probably occurred by chance. In other words, there may be a combination of genetic and nongenetic (i.e., environmental) factors that contributed to the development of cancers within a family. In such instances, where an alteration in a single major gene is not likely or is not identified, individuals may still face elevated risks of cancer.

Deletion or inactivation of tumor suppressor genes can give rise to either familial or sporadic cancer. For example, it was shown in the slide on the evolution of colon cancer that deletions of chromosome regions 18q and 17p, which contain the DCC (deleted in colon carcinoma) and the p53 genes, respectively, are involved in the progression of colon carcinoma. Cancer genetics-©Aayudh Das

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The Rb gene and the p53 gene will be the subjects of further discussion. The next three slides look at the mechanisms by which tumor suppressor genes get mutated to give rise to familial and sporadic cancers.

RETINOBLASTOMA PROTEIN

The retinoblastoma protein (abbreviated pRb, RB or RB1) is a tumor suppressor protein that is dysfunctional in several major cancers. One function of pRb is to prevent excessive cell growth by inhibiting cell cycle progression until a cell is ready to divide. It is also a recruiter of several chromatin remodeling enzymes such as methylases and acetylases.

pRb belongs to the pocket protein family, whose members have a pocket for the functional binding of other proteins. Should an oncogenic protein, such as those produced by cells infected by high-risk types of human papillomaviruses, bind and inactivate pRb, this can lead to cancer. • • • • About 40% retinoblastoma cases are familial It inherits as incompletely penetrant dominant character Familial cases are bilateral, whereas the sporadic forms are unilateral Age-of-onset distribution of bilateral cases is consistent with a single mutation, while sporadic cases followed two-hit kinetics. In 1971Knudson proposed that all retinoblastoma involved two ‘hits’, but that in the familial cases one hit was inherited

The vast majority of cancers are non-hereditary ("sporadic cancers"). Hereditary cancers (familial) are primarily caused by an inherited genetic defect.

Cancer genetics-©Aayudh Das

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Retinoblastoma arises when both copies of the RB gene are inactivated. In the inherited form of the disease, one parental chromosome carries an alteration in this region. A somatic event in retinal cells that causes loss of the other copy of the RB gene causes a tumor. In the sporadic form of the disease, the parental chromosomes are normal, and both RB alleles are lost by (individual) somatic events.

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In 1978 Yunis and Francke showed that retinoblastoma tumor had deletion in chromosome 13q14.

In 1983, recessive nature of the retinoblastoma gene was hypothesized and it was suggested that both alleles of this gene would need to be inactivated for retinoblastoma to arise RFLP analysis of the 13q14 region showed heterozygous state in normal cells of a Rb patient but hemizygous ( only one allele instead of two in a diploid cell) in the tumor tissue of the same individual. This loss of heterozygosity (LOH) suggests that Rb arises through loss of both functional RB alleles Loss of both wild type alleles is seen both in familial and sporadic form of Rb. In the familial form, one germline mutation is inherited, followed by a 2nd somatic event; in the sporadic form both events are somatic.

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Cell-cycle dependent phosphorylation of Rb

Cancer genetics-©Aayudh Das

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Function of the Rb protein

Tumour suppressors may control the cell cycle

As well as occurring in RB itself, mutations are found in the small inhibitory proteins (most notably pi6 and possibly p21), and D cyclin(s). Although these proteins (most notably RB) play a role in the cycle of a proliferating cell, the role that is relevant for tumorigenesis is more probably their function in the quiescent (GO) state. In quiescent cells, RB is not phosphorylated, D cyclin levels are low or absent, and pi6, p21, and p27 ensure inactivity of cdk-cyclin complexes. Cancer genetics-©Aayudh Das Page 4

Chromosomal mechanisms that could lead to loss of function due to loss of heterozygosity (LOH)

Detection of TSG by LOH Analysis

Cancer genetics-©Aayudh Das

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The p53 Functional Circuit

Cancer genetics-©Aayudh Das

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Tumor suppressor p53 suppresses growth or triggers apoptosis

Cancer genetics-©Aayudh Das

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The Mutation Spectrum of p53

p53 is a DNA-binding protein that recognizes an interrupted palindromic 10 bp motif. The ability to bind to its specific target sequences is conferred by the central domain.

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p53 activates transcription at promoters that contain multiple copies of this motif. The immediate N-terminal region provides the transactivator domain. p53 may repress other genes; the mechanism is unknown. p53 also has the ability to bind to damaged DNA. The C-terminal domain recognizes singlestranded regions in DNA.

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p53 is a tetramer (oligomerization is a prerequisite for mutants to behave in a dominant negative manner). Oligomerization requires the C-terminal region. A (putative) signaling domain contains copies of the sequence PXXP, which forms a binding site for SH3 domains.

Cancer genetics-©Aayudh Das

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Cancer genetics-©Aayudh Das

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RB-p53-CDKN2A (INK4A)-p21Network

Cancer genetics-©Aayudh Das

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APC and FAP
• • Intracellular protein Truncations in APC – – • • • Aberrant activation of Wnt pathway

Autosomal dominantly inherited disease Mutations in APC also found in: – – – Sporadic colon cancer

Increased cell proliferation and adenomatous lesions

Hundreds or thousands of polyps in the colon and rectum

Several types of tumours

Wnt signaling and cancer

Hepatocellular carcinoma

Cancer genetics-©Aayudh Das

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Cancer genetics-©Aayudh Das

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Cancer genetics-©Aayudh Das

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