Gene Mutation And Cancer

Md. Mohsin Uddin Howlader 12/22/2012

Assignment On Gene Mutation And Cancer

Submitted To: Dr. Kabirul Bashar Lecturer Department Of Zoology Jahangirnagar University, Dhaka

Submitted by:

Md. Mohsin Uddin Howlader Roll no: 468 Session: 2010-11 Department Of Zoology Jahangirnagar University, Dhaka

Submssion date:

5th January, 2013

Gene Mutation
A gene mutation is a personal change in the DNA sequence that makes up a gene. Mutation range in size from a single DNA building block (DNA base) to a large segment of a chromosome.

Types of gene mutation

The DNA sequence of a gene can be altered a number of ways. Whether they alter the function of essential proteins. The types of mutation include: Missense mutation This type of mutation is a change in one DNA base pair that results in the substitution of one amino acid for another in the protein made by a gene.

Nonsense mutation A nonsense mutation is also a change in one DNA base pair. Instead of substituting one amino acid for another, however the altered DNA sequence prematurely signals the cell to stop building a protein.

Insertion An incertion changes the number of DNA bases in a gene by adding a piece of DNA. As a result the protein made by the gene may not function properly.

Deletion A deletion changes the number of DNA Bases By Removing a piece of DNA.

Duplication A duplication consists of a piece of DNA that is abnormally copied one or more times

Point mutation The replacement base pair by another. This are two types: 1.Transitions: A purine(or a pyrimidine) replaced by another. 2.Transversion Replacement of a by a pyrimidine

Frameshift mutations This occur when one or more base pair are inserted in or detected from DNA, changing the genes leading frame.

Reapeat expansion Nucleotide repeats are short DNA sequence that repeated a number of times in a row. eg-Trinucleotide repeat is made 3 base pair sequences.

Detecting mutations in human genes

In the first decades of medical genetics, both physicians and patients associated this field with uncommon diseases, relevant only to specialists and those affected. This is no longer so: our understanding of the genetics underlying susceptibility to common disorders, such as diabetes mellitus, cardiovascular disease and various cancers, is bringing genetics into not only the medical mainstream, but also the public mainstream. Moreover, molecular methods are becoming important in the diagnosis and classification of malignant disease, as well as in determining prognosis and monitoring response to therapy. This paper describes current methods of studying the changes that underlie inherited and acquired disease and concludes with discussion of a technique that is generating excitement while still in development, the use of DNA microarrays or chips.

How does DNA encode information?

The gene is the fundamental unit of genetic information. Each gene is a DNA sequence tens to millions of nucleotide bases in length and bounded by recognized control regions which produces one or several related proteins. Genetic information is encoded by the sequence of the bases along a single strand of DNA; the bases are therefore often regarded as the alphabet forming the words, phrases and sentences of the genetic instruction manual. When 2 DNA strands combine to form a double helix, the nucleotide bases of one strand associate one-to-

one with those on the other. Genetic information is transmitted by the geometry of pairing of the bases in this double helix. Only certain associations among the 4 different types of bases are permitted, such that there are 2 sets of pairs allowed. As a consequence, either strand alone contains the information required to rebuild the other.

DNA Repair Mechanisms and Human Disease

DNA repair processes exist in both prokaryotic and eukaryotic organisms, and many of the proteins involved have been highly conserved throughout evolution. In fact, cells have evolved a number of mechanisms to detect and repair the various types of damage that can occur to DNA, no matter whether this damage is caused by the environment or by errors in replication. Because DNA is a molecule that plays an active and critical role in cell division, control of DNA repair is closely tied to regulation of the cell cycle. (Recall that cells transit through a cycle involving the G1, S, G2, and M phases, with DNA replication occurring in the S phase and mitosis in the M phase.) During the cell cycle, checkpoint mechanisms ensure that a cell's DNA is intact before permitting DNA replication and cell division to occur. Failures in these checkpoints can lead to an accumulation of damage, which in turn leads to mutations.

Defects in DNA repair underlie a number of human genetic diseases that affect a wide variety of body systems but share a constellation of common traits, most notably a predisposition to cancer. These disorders include ataxia-telangiectasia (AT), a degenerative motor condition caused by failure to repair oxidative damage in the cerebellum, and xeroderma pigmentosum (XP), a condition characterized by sensitivity to sunlight and linked to a defect in an important ultraviolet (UV) damage repair pathway. In addition, a number of genes that have been

implicated in cancer, such as the RAD group, have also been determined to encode proteins critical for DNA damage repair.

Cancer cells
Cancer cells are cells that grow and divide at an unregulated, quickened pace. Although cancer cells can be quite common in a person they are only malignant when the other cells (particularly natural killer cells) fail to recognize and/or destroy them. In the past a common belief was that cancer cells failed to be recognized and destroyed because of a weakness in the immune system. However, more recent research has shown that the failure to recognize cancer cells is caused by the lack of particular co-stimulated molecules that aid in the way antigens react with lymphocytes.

Characteristics of cancer cells

Exhibit Uncontrolled Growth (Immortality)

Abnormal DNA (Mutations) Apoptosis

Cells with damaged DNA that cannot be repaired normally undergo apoptosis, a process in which the cell kills itself. Tumor cells have high levels of a protein (survivin) that inhibits apoptosis.

Lack Differentiation

Normally, cells become differentiated and become capable of specific functions

Lack Contact Inhibition

Normal cells stop dividing when they become crowded because mitosis is inhibited when cells contact nearby cells. Cancer cells continue to divide and produce a mass of cells called a tumor.
Lack Anchorage Dependence

Normal cells cling to neighboring cells, cancer cells do not.

Have a Reduced Need for Growth Factors

Normal cells stop dividing when they become crowded because mitosis is inhibited when cells contact nearby cells. Cancer cells continue to divide and produce a mass of cells called a tumor.
Ability to Penetrate the Lamina

The lamina is a noncellular barrier that is attached to cells that line the surfaces, internal cavities, and organs (epithelial tissue). Normally, cells cannot penetrate this barrier and therefore cannot invade neighboring tissues and organs.

Immune System

The immune system can recognize foreign cells and invaders (bacteria, viruses, etc.) because they have proteins and other structures that are different from the bodys "self" markers.

Angiogenesis

Tumors need a blood supply for food and oxygen. Cancer cells release growth factors that cause nearby blood vessels to produce branches that grow into the cancerous tissue.

Oncogenes

A gene that contributes to the production of a cancer. Oncogenes are generally mutated forms of normal cellular genes (proto-oncogenes). A gene capable, when activated, of transforming a cell. Oncogenes are found in the oncogenically activated state in retroviruses and transformed cells and in their normal non-oncogenically activated state in nontransformed cells in which they are called proto-oncogenes.

Classification
There are several systems for classifying oncogenes, but there is not yet a widely accepted standard. They are sometimes grouped both spatially (moving from outside the cell inwards) and chronologically (parallelling the "normal" process of signal transduction). There are several categories that are commonly used:

Category Growth factors, or mitogens

Examples c-Sis

Cancers glioblastomas, fibrosarcomas, osteosarcomas, breast carcinomas, and melanomas

Gene functions induces cell proliferation.

Receptor tyrosine kinases

epidermal growth factor Breast cancer, receptor (EGFR), platelet-derived growth factor receptor (PDGFR), and vascular endothelial growth factor receptor (VEGFR), HER2/neu gastrointestinal stromal tumours, non-small-cell lung cancer and pancreatic cancer

transduce signals for cell growth and differentiation.

Cytoplasmic tyrosine kinases

Src-family, Syk-ZAP70 family, and BTK family of tyrosine

colorectal and breast cancers, melanomas, ovarian cancers, gastric

mediate the responses to, and the activation

kinases, the Abl gene in CML - Philadelphia chromosome

cancers, head and neck cancers, pancreatice cancer, lung cancer, brain cancers, and blood cancers

receptors of cell proliferation, migration, differentiation, and survival Involved in organism development, cell cycle regulation, cell proliferation, differentiation, cells survival, and apoptosis

Cytoplasmic Serine/threonine kinases and their regulatory subunits

Raf kinase, and cyclindependent kinases (through overexpression).

malignant melanoma, papillary thyroid cancer, colorectal cancer, and ovarian cancer

Regulatory GTPases

Ras protein

adenocarcinomas of the pancreas and colon, thyroid tumors, and myeloid leukemia

involved in signalling a major pathway leading to cell proliferation. -They regulate transcription of genes that induce cell proliferation.

Transcription factors

myc gene

malignant T-cell lymphomas and acute myleoid leukemias, breast cancer, pancreatic cancer, retinoblastoma, and small

Tumer suppressor genes

A tumor suppressor gene, or anti-oncogene, is a gene that protects a cell from one step on the path to cancer. When this gene is mutated to cause a loss or reduction in its function, the cell can progress to cancer, usually in combination with other genetic changes.

nlike oncogenes, tumor suppressor genes generally follow the "two-hit hypothesis", which implies that both alleles that code for a particular protein must be affected before an effect is manifested. This is because if only one allele for the gene is damaged, the second can still produce the correct protein. In other words, mutant tumor suppressors alleles are usually recessive whereas mutant oncogene alleles are typically dominant. The two-hit hypothesis was first proposed by A.G. Knudson for cases of retinoblastoma. Knudson observed that the age of onset of retinoblastoma followed 2nd order kinetics, implying that two independent genetic events were necessary. He recognized that this was consistent with a recessive mutation involving a single gene, but requiring biallelic mutation. Oncogene mutations, in contrast, generally involve a single allele because they are gain-of-function mutations. There are exceptions to the "two-hit" rule for tumor suppressors, such as certain mutations in the p53 gene product. p53 mutations can function as a "dominant negative", meaning that a mutated p53 protein can prevent the function of normal protein from the un-mutated allele. Other tumor-suppressor genes that are exceptions to the "two-hit" rule are those that exhibit haploinsufficiency for example PTCH in medulloblastoma. An example of this is the p27Kip1 cell-cycle inhibitor, in which mutation of a single allele causes increased carcinogen susceptibility.

Functions
Tumor-suppressor genes, or more precisely, the proteins for which they code, either have a dampening or repressive effect on the regulation of the cell cycle or promote apoptosis, and sometimes do both. The functions of tumor-suppressor proteins fall into several categories including the following:

1. Repression of genes that are essential for the continuing of the cell cycle. If these genes are not expressed, the cell cycle does not continue, effectively inhibiting cell division. 2. Coupling the cell cycle to DNA damage. As long as there is damaged DNA in the cell, it should not divide. If the damage can be repaired, the cell cycle can continue. 3. If the damage cannot be repaired, the cell should initiate apoptosis (programmed cell death) to remove the threat it poses for the greater good of the organism. 4. Some proteins involved in cell adhesion prevent tumor cells from dispersing, block loss of contact inhibition, and inhibit metastasis. These proteins are known as metastasis suppressors. 5. DNA repair proteins are usually classified as tumor suppressors as well, as mutations in such their genes increase the risk of cancer, for example mutations in HNPCC, MEN1 and BRCA. Furthermore, increased mutation rate from decreased DNA repair leads to increased inactivation of other tumor suppressors and activation of oncogenes.

Examples
The first tumor-suppressor protein discovered was the Retinoblastoma protein (pRb) in human retinoblastoma; however, recent evidence has also implicated pRb as a tumor-survival factor.

Another important tumor suppressor is the p53 tumor-suppressor protein encoded by the TP53 gene. Homozygous loss of p53 is found in 70% of colon cancers, 3050% of breast cancers, and 50% of lung cancers. Mutated p53 is also involved in the pathophysiology of leukemias, lymphomas, sarcomas, and neurogenic tumors. Abnormalities of the p53 gene can be inherited in Li-Fraumeni syndrome (LFS), which increases the risk of developing various types of cancers.

PTEN acts by opposing the action of PI3K, which is essential for anti-apoptotic, pro-tumorogenic Akt activation.

Other examples of tumor suppressors include VHL, APC, CD95, ST5, YPEL3, ST7, and ST14.

Molecular mechanism of oncogene activation

From proto-oncogene to oncogene

The proto-oncogene can become an oncogene by a relatively small modification of its original function. There are three basic methods of activation:

1. A mutation within a proto-oncogene, or within a regulatory region (for example the promoter region), can cause a change in the protein structure, causing

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an increase in protein (enzyme) activity a loss of regulation

2. An increase in the amount of a certain protein (protein concentration), caused by

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an increase of protein expression (through misregulation) an increase of protein (mRNA) stability, prolonging its existence and thus its activity in the cell

gene duplication (one type of chromosome abnormality), resulting in an increased amount of protein in the cell

3. A chromosomal translocation (another type of chromosome abnormality)

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There are 2 different types of chromosomal translocations that can occur: 2. translocation events which relocate a proto-oncogene to a new chromosomal site that leads to higher expression 3. translocation events that lead to a fusion between a proto-oncogene and a 2nd gene (this creates a fusion protein with increased cancerous/oncogenic activity)

the expression of a constitutively active hybrid protein. This type of mutation in a dividing stem cell in the bone marrow leads to adult leukemia

Philadelphia Chromosome is an example of this type of translocation event. This chromosome was discovered in 1960 by Peter Nowell and David Hungerford, and it is a fusion of parts of DNA from chromosome 22 and chromosome 9. The broken end of chromosome 22 contains the "BCR" gene, which fuses with a fragment of chromosome 9 that contains the "ABL1" gene. When these two chromosome fragments fuse the genes also fuse

creating a new gene: "BRC-ABL". This fused gene encodes for a protein that displays high protein tyrosine kinase activity (this activity is due to the "ABL1" half of the protein). The unregulated expression of this protein activates other proteins that are involved in cell cycle and cell division which can cause a cell to grow and divide uncontrollably (the cell becomes cancerous). As a result, the Philadelphia Chromosome is associated with Chronic

Myelogenous Leukemia (as mentioned before) as well as other forms of Leukemia.

The expression of oncogenes can be regulated by microRNAs (miRNAs), small RNAs 21-25 nucleotides in length that control gene expression by downregulating them. Mutations in such microRNAs (known as oncomirs) can lead to activation of oncogenes. Antisense messenger RNAs could theoretically be used to block the effects of oncogenes.