CANCER GENETICS

Dr. Zeyad Akawi Jreisat, M.D., M.A., Ph.D.

CANCER
Cancer is a genetic disease because it can be traced to alterations within specific genes, but in most cases, it is not an inherited disease

Initiation
Irreversible with memory Requires fixation by cell division Additive No readily measurable threshold For many chemical initiators, dependent upon xenobiotic metabolizing capabilities of the cell Single initiated cells are not usually identifiable

Promotion
Reversible Promoted cell population dependent upon continued presence of the promoting agent for their existence Dose response exhibits a threshold and maximal effect level Modulated by variety of environmental factors, including age, diet, hormonal status, and frequency of administration of promoting agent Although not carcinogenic, promoting agents may promote fortuitously initiated cells Promoted lesions are seen microscopically and/or grossly

Progression
Irreversibility Growth of altered cells responsive to environmental factors during early phase of progression Discernable alterations in cell genome Evolving chromosomal abnormality Fortuitous progression of promoted cells can be demonstrated Benign and/or malignant neoplasm seen

CANCER
Genes play an important role in cancer Cancer can be viewed as a series of genetic changes that eventually lead to uncontrolled cell growth Cancer usually is a multi-step process A cancerous growth can be considered clonal in origin 5-10% of cancer patients have an inherited predisposition to develop the cancer Most cancers 90-95% are not passed from parents to offspring Once a cellular growth has became cancerous or malignant, the cancer cells are invasive and metastatic

1. DNA of a normal cell

This piece of DNA is an exact copy of the DNA from which it came. When the parent cell divided to create two cells, the cell's DNA also divided, creating two identical copies of the original DNA.

2. Mutation of DNA

This DNA has suffered a mutation, either through miscopying (when its parent cell divided), or through the damaging effects of exposure to radiation or a chemical carcinogen.

3. Genetically altered cell

The DNA of the cell highlighted above has a mutation that causes the cell to replicate even though this tissue doesn't need replacement cells at this time or at this place.

4. Spread and second mutation

After about a million divisions, there's a good chance that one of the new cells will have mutated further. This cell, now carrying two mutant genes, could have an altered appearance and be even more prone to reproduce unchecked.

5. Third mutation

Over time and after many cell divisions, a third mutation may arise. If the mutation gives the cell some further advantage, that cell will grow more vigorously than its predecessors and thus speed up the growth of the tumour.

6. Fourth mutation

The new type of cells grow rapidly, allowing for more opportunities for mutations. The next mutation paves the way for the development of an even more aggressive cancer. At this point the tumour is still contained.

7. Breaking through the membrane

The newer, wilder cells created by another mutation are able to push their way through the epithelial tissue's basement membrane, which is a meshwork of protein that normally creates a barrier. The invasive cells in this tumour are no longer contained. At this point the cancer is still too small to be detected.

8. Angiogenesis

Often during the development of earlier stages of the tumour, or perhaps by the time the tumour has broken through the basement membrane (as pictured above), angiogenesis takes place. Angiogenesis is the recruitment of blood vessels from the network of neighbouring vessels.

9.Invasion and dispersal

The tumour has now invaded the tissue beyond the basement membrane.

The Most Common Cancer Risk Factors

Genetic predisposition: Colon and Breast cancer. High estrogen exposure: Breast and uterine cancer.

Ionizing radiation: X-Rays and nuclear radiation.

Ultraviolet radiation: Skin cancer. Carcinogenic chemicals: Asbestos, Polycyclic aromatic hydrocarbons, Aromatic amines, Nitrosamines, Various drugs, Inorganic compounds

 Tobacco smoke: Lung cancer

Alcohol: Mouth, throat, esophagus, stomach, and liver cancer Carcinogenic foods: Include salted, and smoked foods, meats treated with nitrites

Free radicals: Highly reactive chemical compounds that can damage DNA
Unhealthy diet: High in saturated fat (colon, rectum and prostate gland cancer)

 Viruses: DNA viruses: Papovavirus (papilomavirus, polyomavirus, SV40), Adenovirus, Herpesvirus, Hepadna virus family RNA viruses: Retrovirus type C, Retrovirus type B Examples: - Epstein-Bar virus associated with Burkitts lymphoma, nasopharyngeal carcinoma, B cell lymphomas - Hepatitis B virus associated with hepatocellular carcinoma - Human papillomaviruses associated with anogenital cancers

AMES TEST
This test used to screen for potential carcinogens using a specially constructed strain of Salmonella typhimurium that has a mutation (His-) in a gene that codes for one of the enzymes involved in the synthesis of histidine. Salmonella typhimurium + Histidine Growth Salmonella typhimurium - Histidine No Growth Salmonella typhimurium + Carcinogens Mutations His- His+ Salmonella typhimurium - Histidine Growth

DNA is the critical macromolecule in carcinogenesis WHY ?

The essential changes responsible for cancer are transmitted from mother to daughter cells Both irradiation and chemical carcinogens damage DNA and are capable of causing mutations in DNA A high incidence of cancer in individuals who have an inherited deficiency in their ability to repair lesions in DNA Many tumor cells exhibit abnormal chromosomes Transfection experiments indicate that purified DNA (oncogenes) from cancer cells can transform normal cells into cancer cells The presence of special genes in the cells showed to increase susceptibility to cancer

Changes shown by cultured cells after being transformed into malignant cells
Morphology changes Increased cell density Loss of anchorage dependence Loss of contact inhibition Biochemical changes Alteration of cytoskeletal structure Diminished requirement for growth factors

Cancer Causing Genes Usually involved In

The progression of a cell through the cell cycle.
Adhesion of a cell to its neighbors. Apoptosis. DNA repair.

Types of genes which may mutate to cause cancer

Tumour suppressor genes Oncogenes DNA repair genes Telomerases

Tumour Suppressor Genes

Tumor Suppressor Genes

Mutations in both alleles or a mutation in one followed by a loss of or reduction to homozygosity in the second Loss of function of a protein Mutation present in germ cell (can be inherited) or somatic cell Strong tissue preference (effect of RB1 gene in the retina)

Knudsens two hit hypothesis

Proteins which are encoded by tumor-suppressor genes

1 Intracellular proteins that regulate or inhibit progression through a specific stage of the cell cycle (e.g., p16 and Rb). 2 Receptors or signal transducers for secreted hormones or developmental signals that inhibit cell proliferation (e.g., TGF). 3 Checkpoint-control proteins that arrest the cell cycle if DNA is damaged or chromosomes are abnormal (e.g., p53). 4 Proteins that promote apoptosis. 5 Enzymes that participate in DNA repair.

Mechanisms for inactivating tumorsuppressor genes

1) Have deletions, point mutations or gene conversion that prevent production of any protein or lead to production of a non-functional protein. 2) Methylation of cytosine residues in the promoter or other control elements. Such methylation is commonly found in non-transcribed regions of DNA. 3) Chromosomal abnormalities

Mechanisms of loosing the remaining good copy of a tumor suppressor gene

Tumor suppressor genes mutations Causing Loss of Growth-Inhibition and Cell-Cycle Controls

p15: protein which causes cells to arrest in G1.

Binding of TGF to its receptor induces activation of cytosolic Smad transcription factors After translocating to the nucleus, Smads can promote expression of the gene encoding p15.

p16: binds specifically to CDK4 and CDK6, cyclindependent kinases (CDKs), thereby inhibiting their kinase activity (whose promotes progression past the restriction point) and causing G1 arrest. the p16 gene is inactivated by hypermethylation of its promoter region, which Prevents transcription.

Cyclin D1 :Amplification of the cyclin D1 gene and

concomitant overproduction of the cyclin D1 protein is common in human breast cancer.

Rb: The complete phosphorylation of Rb irreversibly commits

the cell to DNA synthesis. phosphorylation of Rb protein is initiated midway through G1 by active cyclin D-CDK4 and cyclin D-CDK6 complexes & completed by other cyclin-CDK complexes in late G1, allowing activation of E2F transcription factors, which stimulate transcription of genes encoding proteins required for DNA synthesis.
The kinase activity of cyclinD-CDk phosphorylates Rb, thereby activating E2F; this kinase activity is inhibited by p16. In human cancers either a positive regulator overproduction of cyclin D, or negative regulators p16 and Rb. Rb function can be eliminated by the binding of an inhibitory protein, designated E7, that is encoded by human papillomavirus (HPV)

Interaction between the Rb and E2F protein

p53:
Is a sensor essential for the checkpoint control that arrests cells with damaged DNA in G1. Unlike other cell-cycle proteins, p53 is present at very low levels in normal cells because it is extremely unstable and rapidly degraded. DNA damage by -irradiation or by other stresses somehow leads to the activation of ATM, a serine kinase that phosphorylates and thereby stabilizes p53, leading to a marked increase in its concentration. The stabilized p53 activates expression of the gene encoding p21 CIP, which binds to and inhibits mammalian G1 cyclin-CDK complexes.

 Oncogenic p53 mutations act as dominant negatives, with mutations in a single allele causing a loss of function. The activity of p53 normally is kept low by a protein called Mdm2. Mdm2 gene is itself transcriptionally activated by p53, Mdm2 functions in an autoregulatory feedback loop with p53, perhaps normally preventing excess p53 function. The activity of p53 also is inhibited by a human papillo-mavirus (HPV) protein called E6.

Central role of p53 in preventing the proliferation of cancer cells

APC protein: inhibits the progression of certain types of cells through the cell cycle & it does so by preventing the Wnt signal-transduction pathway from activating expression of protooncogenes including the c-myc gene. The first step in colon carcinogenesis involves loss of a functional APC gene, resulting in formation of polyps (precancerous growths)