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BIOCHEMISTRY OF CANCER
1.
2.
3.
4.
5.
6.

Write a note on agents which bring about carcinogenesis. Pon Nov 2010
Explain the mechanisms involved in carcinogenesis. Pon May 2010
Oncogenes. MGR Aug 2008
Mutagens MGR Feb 2012
Carcinogenic virus. MGR Aug 2009
Name two tumour suppressor genes and the malignancy that is specifically
associated with abnormalities in each of these genes. MGR Feb 2014
7. Write a note on tumor suppressor genes. Pon Nov 2010
8. Carcinogenic virus.
ETIOLOGY OF CANCER:
1. All cancers originate from one aberrant cell. During surveillance by the immune
system, these aberrant cells are usually destroyed. As age advances, the probability of
the incidence of cancer is increased.
2. Cancers are multifactorial in origin. They include genetic, hormonal, metabolic,
physical, chemical and environmental factors.
MUTAGENS:
1. Any substance which increases the rate of mutation can also enhance the rate of
incidence of cancer. Therefore all carcinogens are mutogens.
2. Examples are Chemicals, X-ray, gamma-ray, ultraviolet ray etc.
CHEMICAL CARCINOGENS:
Classification:
1. Organic
a. Dimethyl benzanthracene,
b. Benzo pyrene,
c. Dimethyl nitrosamine
2. Inorganic
a. Arsenic,
b. Cadmium
3. Occupation: Asbestos, benzene.
4. Diet: Aflatoxin B
5. Drugs:
a. Hormones: diethylstibesterol.
b. Chemotherapeutic agents
6. Life style: cigarette smoking.
Mechanism of action:
1. Chemical carcinogens act cumulatively. They bind to purines, pyrimidines and
phosphodiesterase bonds of DNA causing unrepairable damage. The chemical
carcinogens frequently cause mutations of DNA which may finally lead to the
development of cancer, hence they are regarded as mutagens.
2. Ames assay: This is a laboratory test to check the carcinogenecity of chemicals.
Mutant strain of Salmonella typhimurium can not synthesize histidine. Addition of
chemical carcinogens causes reverse mutation restoring the ability of the bacteria to
synthesize histidine. By detecting this reverse mutation in Salmonella in the colonies
of agar plates, the chemical mutagens can be identified.
3. Promoters: Most carcinogens require promoters for the production of a cancer.
Benzopyrene applied on skin does not produce cancer. Croton oil application also does
not lead to skin cancer. But when benzopyrene application is followed by croton oil,
tumor is developed. In this case, croton oil is termed as the promoter.

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4. Progression: Development of familial adenomatous polyposis is a good example for
multistep progression. Mutations in the APC gene are inherited from parents. By the
time the patient becomes adult, there will progress to form adenomas. Further
mutation lead to the development of malignancy.
5. Chemical carcinogens may produce the cancer: (a) At the sit of exposure, e.g. buccal
cancer in tobacco chewers, skin cancer in tar workers. (b) At the site of metabolism,
e.g. liver cancer produced by aflatoxin. (c) At the site of elimination, e.g. bladder
cancer in persons working with aromatic dyes.
Important chemical carcinogens:
1. Aflatoxins: They are a group of chemically related compounds synthesised by the
fungi, Aspergillus flavus. The mould grows on rice, wheat and groundnut, when kept in
damp conditions. The fungi may grow in cattle fodder, which may enter into human
body through the cow's milk. Aflatoxins are powerful carcinogens, which produce
hepatomas .
2. Cigarette: Lung cancer is associated with the habit of cigarette smoking. Cigarette
contains many carcinogens, the most important group being benzo(a)pyrenes. Other
important deleterious substances in cigarette smoke are nicotine, carbon monoxide,
nitrogen dioxide and carbon soot.
3. Oral cancer is strongly associated with chewing of tobacco. Oral cancer constitutes
20% of all cancers seen in India, whereas it is less than 1% in Western countries.
4. Alcohol intake increases the risk of oral, pharyngeal, esophageal and liver cancers.
Diet high in total fat and cholesterol increases the risk of colon, breast and prostate
cancers.
Action of Chemical Carcinogens
1. Chemical carcinogens are ingested as procarcinogens. The enzymes responsible for
the activation of procarcinogens are cytochrome P-450 system .
2. On the other hand, direct carcinogens are the ones which interact directly with the
target molecules, e.g. methyl cholanthrene.
PHYSICAL CARCINOGENS
X-ray, gamma-ray and UV-ray may cause:
1. Formation of pyrimidine dimers,
2. Apurinic sites with consequent break in dna, and
3. Formation of free radicals and superoxides which cause DNA break, leading to somatic
mutations.
4. Exposure of X-ray in fetal life will increase the risk of leukemia in childhood. In
population studies, 1 rad per year will increase the cancer incidence by 40/ million
people per year.
Antimutagens
1. These are substances which will interfere with tumor promotion. Vitamin A and
carotenoids are shown to reverse precancerous conditions.
2. Vitamin E acts as an antioxidant, preventing the damage made by free radicals and
superoxides.
3. Vitamin C regularly given to persons working with aniline prevented the production
of new cancer cases.
4. Tubers, beans and leafy vegetables are shown to interrupt tumor promotion.
5. Curcumin, the yellow substance in Turmeric is known to prevent mutations.
6. Fiber content of the diet is effective in colon cancers.
7. Low protein, low fat, diet decreases the risk of cancer in animal studies.
ONCOGENIC VIRUSES
1. Viruses involved in the development of cancer are known as oncogenic viruses which

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may contain either DNA or RNA.
2. Viral genes enter the cell and gets integration into the host DNA. The drive for
multiplication by the virus genome overrules the regulatory checks of the cellular
mechanism. So, there is uncontrolled multiplication of the cells. This is called
transformation by oncogenic virus.
3. Human oncogenic viruses:
Virus
Epstein-Barr
virus
Human
papilloma virus
Hepatitis B
virus

Abbreviati
on
EBV

Associated human cancer

HPV

Burkitt's lymphoma (BL); Nasopharyngeal

carcinoma (NPC)
Uterine cervical carcinoma

HBV

Hepatoma

ONCOGENES:
1. The viral genes capable of causing cancer are originally known as oncogene. Normal cells
also contain DNA sequences similar to viral oncogenes. These genes are called
protooncogenes.
2. Proto-oncogenes control normal cell growth and division.
3. Examples of proto-oncogenes:
a. Growth factors,
b. Growth factor receptors,
c. Signal transduction proteins,
d. Transcription factors,
e. Cell cycle regulators, and
f. Regulators of apoptosis.
4. The mutations in above proto oncogenes can give rise to oncogenes, an important step in
the causation of cancer.
5. Mechanisms of converting the proto-oncogenes to oncogenes:
a. Radiation and chemical carcinogens act by producing a mutation in the coding
portion of the prto-oncogene converting into onccogenes.
b. Viral insertionin to chromosome:
i. When certain retroviruse (genetic materia RNA) infect cells, a
complementary DNA (cDNA) is made from their RNA by the enzyme reverse
transcriptase. The cDNA so produced gets inserted into the host genome.
This pro-viral DNA takes over the control of the transcription of cellular
chromosomal DNA
ii. Some DNA viruses also get inserted in to the host chromosome and activate
the protooncogenes.
c. Chromosomal translocation resulting in overexpression of proto-oncogenes. Eg.
Burkitt's lymphoma.
d. Gene amplification: Some oncogenes result when multiple copies of a protooncogene are created. This occurs in certain drug administration like methotrexate
and may cause resistance to cancer therapy.
e. Point mutation: It refers to a change in a single base in the DNA. The ras
protooncogene is an example of activation by point mutation. The mutated ras
oncogene produces a protein (GTPase) which differs in structure by a single amino
acid. This alteration diminishes the activity of GTPase, involved in the control of
cell growth.

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6. Mechanism of action of oncogenes:
Oncogenes encode for oncoproteins. These proteins are involved in the transformation
and multiplication of cells.
7. Oncoprotenes:
a. Growth factors: Normally the cell proliferation is stimulated by growth factors.
Overexpression and/or structural alterations in growth factor receptors are
associated with carcinogenesis. Transforming growth factor (TGF-d) is a protein
synthesized and required for the growth of epithelial cells. TGF-a is produced in
high concentration in individuals suffering from psoriasis.
b. Growth factor receptors: Some genes encoding growth factor receptors have
been identified to become oncogenes. Eg.The overexpression of gene erb-9,
encoding ECF receptor is observed in lung cancer.
a. GTP-binding proteins: These are a group of signal transducing proteins.
Guanosine triphosphate (GTP)-binding proteins are found in about 30%" of human
cancers. The mutation of ras protooncogene is the cause of many human tumors.
For Eg. Point mutations in ras gene result in lack of GTPase activity. This leads to
the occurrence of ras P21 in a permanently activated state, causing uncontrolled
multiplication of cells.
b. Non-receptor tyrosine kinases : These proteins are found on the interior of the
inner plasma membrane. They phosphorylate the cellular proteins involved in cell
division in response to external growth stimuli. Mutations in the
protooncogenes(eg. abl) encoding nonreceptor tyrosine kinases increase the
kinase activity and, in turn, phosphorylation of target proteins causing unlimited
cell multiplication.
8. Antioncogcnes
Cancer suppressor genes or anti oncogenes apply breaks and regulate cell
proliferation. The loss of these suppressor genes removes the growth control of cells
e.g. retinoblastoma, one type of breast cancer, carcinoma of lung, Wilms' kidney
tumor.
9. Genes that regulate apoptosis:
Genes that regulate programmed cell death (apoptosis) are important in the
development of tumors. Gene bcl-2 causes B-cell lymphoma by preventing
programmed cell death. Over expression of bcl-2 allows other mutations of
protooncogenes that, ultimately, leads to cancer.

TUMOUR SUPPRESSOR GENES:

1. Tumor suppressor genes (anti oncogenes) are normal genes that slow down cell
division, repair DNA mistakes, or initiate programmed cell death. If they don't work
properly, it can lead to cancer.
2. They become abnormal if both copies (alleles) of the gene are inactivated ("two-hit
hypothesis). Thus, mutant tumor suppressors' alleles are usually recessive whereas
mutant oncogene alleles are dominant. Oncogene mutations, in contrast, involve a
single allele because they are gain-of-function mutations.
3. Inherited abnormalities of tumor suppressor genes have been found in some cancer
syndromes. But most tumor suppressor gene mutations are acquired.
4. The two best understood cell cycle regulators that are also tumor suppressors are the
retinoblastoma (rb ) and p53 genes.
5. p53 gene
1. The gene:

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i. Short arm of chromosome 17 contain an oncosuppressor gene, called
p53. It is so called because the gene encodes a phosphoprotein with
molecular weight 53,000;
ii. It acts as the guardian of the genome
iii. Most tumors have a complete absence of p53, whereas others show
mutant nonfunctional p53.
2. Functions of p53:
i. In response to DNA-damaging mutagens, ionizing radiation, or UV light,
the level of p53 rises. p53, acting as a transcription factor, stimulates
transcription of p21
ii. The p21 gene product inhibits the cyclinCDK complexes, which
prevents the phosphorylation of Rb and release of E2F proteins. The cell
is thus prevented from entering S phase.
iii. p53 also stimulates the transcription of a number of DNA repair
enzymes.
iv. If the DNA is successfully repaired, p53 induces its own downregulation
through the activation of the mdm2 gene. If the DNA repair was not
successful, p53 activates several genes involved in apoptosis and
blocking the growth factors.
5. Clinical significance:
i. People who inherit only one functional copy of p53 will most likely
develop tumors in early adulthood, a disease known as Li-Fraumeni
syndrome.
ii. p53 can also be damaged in cells by mutagens (chemicals, radiation or
viruses), increasing the likelihood that the cell will begin uncontrolled
division.
iii. In-vitro introduction of p53 in to p53-deficient cells has been shown to
cause rapid death of cancer cells or prevention of further division.
Restoring its function would be a major step in curing many cancers
Role of telomerase in carcinogenesis. Pon May 2009
1. The ends of each chromosome contain structures called telomeres. Telomeres consist
of short TG-rich repeats of the sequence 5'-TTAGGG-3'.
2. They maintain the structural integrity of the chromosome, preventing attack by
nucleases, and allow repair systems to distinguish a true end from a break in dsDNA.
3. During DNA replication, when it approaches the end of the chromosome, normally a
problem develops in the lagging strand. Either primase cannot lay down a primer at
the very end of the chromosome or, after DNA replication is complete, the RNA at the
end of the chromosome is degraded. Consequently, in most normal human somatic
cells, telomeres shorten with each successive cell division. Once telomeres are
shortened beyond some critical length, the cell is no longer able to divide and is said
to be senescent.
4. Telomerase is a multi subunit RNA-containing enzyme complex is responsible for
telomere synthesis and thus for maintaining the length of the telomere.
5. Telomerase contains a protein that acts as a reverse transcriptase, and a short piece
of RNA that acts as a template.
6. Normally, in germ cells and other stem cells, telomeres do not shorten and the cells do
not senesce. In somatic cells the enzyme is not expressed and senescence and cell
division will not proceed.
7. But in cancer cells, the enzyme is expressed so that they keep multiplying.

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8. Since telomere shortening has been associated with both malignant transformation
and aging, telomerase has become an attractive target for cancer chemotherapy and
drug development.
TUMOR MARKERS
1. What are tumor markers? Pon May 2009
2. Mention two tumour markers and specify the diagnostic application MGR Aug
2011
3. Tumor markers.
Thay are also called as tumor index substances. They are factors released from the
tumor cells, which could be detected in blood and therefore indicate the presence of
the tumor in the body.
Clinically Important Tumor Markers
1. Alpha Fetoprotein (AFP):
1. It is fetal albumin and has similarities with adult albumin. It is increased in the
circulation of patients with hepatocellular carcinoma, germ cell tumors,
teratocarcinoma of ovary and in pregnancy with fetal malformations of neural tube .
2. In adult males and nonpregnant females, normal value is less than 15 ng/L. A value of
AFP above 300 ng/L is often associated with cancer, although levels in this range may
be seen in nonmalignant liver diseases. Levels above 1000 ng/L are almost always
associated with cancer (except in pregnancy). The gene for AFP is located in
chromosome No. 4.
2. Carcinoembryonic Antigen (CEA)
1. CEA level is markedly increased in colorectal cancers. Over 50% of persons with
breast, colon, lung, gastric, ovarian, pancreatic, and uterine cancer have elevated
levels of CEA.
2. CEA levels may also be elevated in inflammatory bowel disease (IBD), pancreatitis,
and liver disease.
3. Heavy smokers and about 5% of healthy persons have elevated plasma levels of CEA.
3. Beta Chain of Chorionic Gonadotropin
1. Beta-HCG is synthesised by normal syncytio trophoblasts (cells of placental villi). HCG
is a glycoprotein; it has alpha and beta subunits. The alpha subunit is identical with
those of FSH, TSH and LH.
2. The beta subunit is specific for HCG. It is increased in hydatidiform mole,
choriocarcinoma and germ cell tumors. About 60% of testicular cancers secrete. hCG.
Normal value is less than 20 IU/L for males and non-pregnant females. Greater than
100,00 IU/ L indicates trophoblastic tumor.
4. Cancer Antigen 125 (CA-125)
1. CA-125 is a tumor maker for ovarian cancers. It is a glycoprotein with a molecular
weight of 10 million; one of the biggest molecules identified. The name is so given
because it reacted with a monoclonal antibody, originally termed as OC-125.
2. Approximately 75% of persons with ovarian cancer will have elevated serum levels.
3. CA-125 levels correlate with tumor mass; consequently, this test is used to determine
whether recurrence of the cancer has occurred following chemotherapy.
4. Normal blood level of CA125 is lessthan 35 U/mL.
5. Tissue Polypeptide Antigen (TPA)
1. It is a common human carcinoma antigen, produced during G2 phase and released
into surrounding fluids during mitosis.
2. It is not specific for cancer of a particular site; but it is useful to assess the activity of

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the tumor. It is seen in blood as long as the tumor cells proliferate.
3. The TPA blood test is sometimes used along with other tumor markers to help follow
patients being treated for lung, bladder, and many other cancers.
6. Prostate Specific Antigen (PSA)
1. It is produced by secretory epithelium of prostate gland. It is normally secreted into
seminal fluid, where it is necessary for the liquefaction of seminal coagulum. It is a
glycoprotein. It is a protease, and in serum it is seen complexed with alpha-1antitrypsin.
2. The PSA level, especially the complexed form, is increased in prostate cancers. PSA
has been found to be elevated in 60-70% patients with cancer of the prostate. Most
PSA is bound to antitrypsins in plasma but some PSA circulates unbound to protein
(free PSA).
3. Normal blood level of total PSA is less than 4 ng/L. Persons with a borderline total PSA
(between 4-10 ng/L), but who have a low free PSA are more likely to have malignant
prostate disease.
Other Tumor Markers used occasionally:
1. Estrogen Receptor (ER)
a. ER is a protein found in the nucleus of breast and uterine tissues.
b. The level of ER in the tissue is used to determine whether a person with breast
cancer is likely to respond to therapy with tamoxifen, which binds to the
receptors blocking the action of estrogen.
c. Women who are ER-negative have a greater risk of recurrence than women who
are ER-positive.
2. Progesterone Receptor (PR)
a. PR consists of two proteins, which are located in the nuclei of both breast and
uterine tissues. PR has the same prognostic value as ER, and is measured by
similar methods. Tissue that does not express the PR receptors is less likely to
bind estrogen analogs used to treat the tumor. Persons who test negative for
both ER and PR have less than a 5% chance of responding to endocrine
therapy.
3. Nuclear Matrix Protein (NMP22)
a. NMP22 is a structural nuclear protein that is released into the urine when
bladder carcinoma cells die. Approximately 70% of bladder carcinomas are
positive for NMP22. Bladder Tumor-associated Analytes (BTA) BTA is comprised
of type IV collagen, fibronectin, laminin, and proteoglycan, which are
components of the basement membrane that are released into the urine when
bladder tumor cells attach to the basement membrane of the bladder wall.
These products can be detected in urine using a mixture ofantibodies to the
four components. BTA is elevated in about 30% of persons with low-grade
bladder tumors and over 60% of persons with high-grade tumors.
4. Beta-2-Microglobulin (B2M)
a. B2M blood levels are elevated in multiple myeloma, chronic lymphocytic
leukemia (CLL), and some lymphomas.
b. B2M is useful to help predict the long-term prognosis in these cancers. Patients
with higher levels of B2M usually have a poorer prognosis. B2M is also checked
during treatment of multiple myeloma to see how well the treatment is
working.
5. Bladder Tumor Antigen (BTA)
a. BTA is found in the urine of many patients with bladder cancer. It may be seen
in some non-cancerous conditions such as kidney stones or urinary tract

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infections.
b. It is sometimes used along with NMP22 to test patients for the recurrence of
bladder cancer. This test is not often used. levels of CA-125 are also found in
approximately 20% of persons with pancreatic and digestive tract.
6. Calcitonin:
a. In medullary thyroid carcinoma (MTC), a cancer of parafollicular C cells, blood
levels of calcitonin are often greater than 100 pg/ml. This is one of the rare
tumor markers that can be used to help detect early cancer.
7. Neuron Specific Enolase (NSE)
a. NSE is a marker for neuroendocrine tumors such as small cell lung cancer,
neuroblastoma, and carcinoid tumors.
8. Thyroglobulin
a. Thyroglobulin is a protein synthesised by the thyroid gland. Thyroglobulin levels
are elevated in many thyroid diseases, including some common forms of
thyroid cancer.