CANCER GENETICS

SREEKUTTY S
2ND MSc ZOOLOGY
 CANCER
• Unregulated proliferation of cell.
• Cause malignant tumour / Neoplasms.
• Metastasis – form 20 tumours.
• Result from genetic malfunctions - triggered by Carcinogens.
• Cells that initiate tumor are not mature cells, but precursor cells - So
mutation is passed on to many progeny cells.
• Cancer cell did not require an external signal for proliferation.
• Fail to sense signals that restrict cell division.
• Loss contact inhibition.
• Cancer cells are immortal.
• It is the Tumour-initiating cells or cancer stem cells (CSCs), is capable of giving
rise to and maintaining tumours.

• The two most commonly used surface markers used to identify CSCs are CD133
and CD44.

• CSC must have ability for self renewal.

-> Self-renewal maintains a reservoir of CSCs when the CSCs undergoes either
asymmetrical or symmetrical division .
-> Asymmetric division forms one more differentiated cell and one CSC.
-> Symmetrical division results in the CSC forming either two differentiated cells
or two CSCs.

• Not only must a CSC be able to self-renew, but it must also be able to differentiate
to recapitulate the heterogeneity seen in tumours.
 ANGIOGENESIS
• Most tumors induce formation of new blood vessels that invade the tumor and
nourish it .
• Tumor producing growth factors that induce angiogenesis are,
-> Basic fibroblast growth factors (BFGF)
-> Transforming growth factor α (TGFα)
-> Vascular endothelial growth factor (VEGF)

ANGIOGENESIS INCLUDES
• Degradation of basal lamina that surround a near by capillary.
• Migration of endothelial cells lining the capillary into the tumor.
• Division of endothelial cells and formation of new basement membrane around
the newly elongated capillary.
BASIS OF OCCURRENCE OF
CANCER
1) CLONAL INHERITENCE

2) ONCOVIRUS

3) INHERITED IN FAMILIES
Show simple dominance , incomplete penetrance , variable expressivity….

4) MUTATION BY CARCINOGENS

PHYSICAL CARCINOGENS
-> Ionizing radiations - X-rays & Gamma rays…
-> Non- ionizing radiations - UV rays, microwave…

CHEMICAL CARCINOGENS
-> Benzopyrene -> Nitrosamine -> Vinyl chloride
-> Aflatoxin -> MSG -> Acrylamide
 CANCER & CELL CYCLE
• Phases of cell cycle ( G, S, M ) & their duration is controlled by external &
internal chemical signals.

• Transition b/w different phases are regulated at checkpoints.

• Checkpoint : Mechanism that halts progression through the cycle until a

critical process , such as DNA synthesis is completed or until damaged DNA
is repaired.

• Checkpoints are regulated by activity of proteins : Cyclin & CDK

• Complexes formed by Cyclin-CDK cause the cell cycle to progress.

In tumour cells, checkpoints are deregulated – due to genetic defects in machinery
that alternately rise / lower abundance of CDK-Cyclin complexes.

: Genes encoding the cyclins or CDK may be mutated.

: Genes encoding proteins that respond to Cyclin-CDK complexes are mutated.

: Genes that regulate abundance of these proteins are mutated.

 MUTATION OF ONCOGENES INCLUDES :

POINT MUTATION DNA REARRANGEMENT

• Single base-pair substitution in • Either deletion or base sequence
DNA. Eg : Seen in C-ras exchange.

TRANSLOCATION GENE AMPLIFICATION

• The c-myc proto-oncogene is • Increase the number of copy of a
translocated from chromosome 8 particular proto oncogene cause over
to the immunoglobulin heavy- production of normal protein rather than
chain locus (IgH) on chromosome formation of abnormal protein.
14 in Burkitt's lymphomas,
• Eg – certain lung cancer caused by the
resulting in abnormal c-
over production Myc protein from C- myc.
myc expression.
 ONCOVIRUS & ONCOGENES
• Oncovirus : Cancer inducing virus.
• They are mainly retroviruses.

• Rous sarcoma virus : 1st tumour inducing virus discovered in 1910 by Peyton Rous.
: Cause sarcoma in connective tissues in chicken.
• They have 4 genes in their RNA
-> gag : Code for capsid
-> pol : Code for enzyme reverse transcriptase.
-> env : Code for proteins in viral envelope.
-> v-src : Code for protein kinases
: It is the only tumour inducing / onco gene in Rous sarcoma virus.
21 different viral oncogenes are discovered yet :

ONCOGENES FUNCTION
abl,, fes, fgr, fps, ros, src, yes Tyrosine specific protein kinase.
rel, myc, myb Transcription factor
raf, mos, mil Serine / threonine protein kinase
H-ras, K-ras GTP – binding protein
jun, fos Transcriptional activator protein
erbB Truncated version of EGF receptor
fms Analogue of CSF-1 receptor
erbA Analogue of TH receptor
sis Analogue of Platelet derived growth
factor (PDGF)
Genes responsible for oncogenesis are,

• Proto-oncogenes

• Tumour suppressor genes

 PROTO-ONCOGENES
• They are cellular homologues of viral oncogenes.

• Also called as normal cellular oncogene – denoted by c-onc.

• In normal conditions products of c-onc play key role in

regulating cellular activities.

• A mutation in these genes can upset the biochemical balance

within a cell - & put it on a track to become cancerous.

• Such mutation transform normal c-onc to onc (oncogene).

 1) Src

• Eg : In tumour cells of chicken, the src

gene produce 100 times as much
tyrosine kinase than c-src.

• This upsets signal mechanisms that

control cell division – causing
unregulated growth.
 2) H-ras

• Robert winberg & colleagues isolated mutation responsible for

human bladder cancer using transfection test.

• Here proto oncogene C-H-ras is responsible.

• Ras-regulated signal pathways control -

actin cytoskeletal integrity, cell proliferation, cell
differentiation, cell adhesion, apoptosis…

• Nucleotide in codon 12 of this allele had been mutated with a

substitution of a valine for glycine.

• Mutant ras is switch to its active signalling mode – It

continuously hydrolase GTP – Enables cells to divide in an
uncontrolled way.
 3) HER-2 / NEU
• Proto oncogene encodes a transmembrane tyrosine
kinase receptor with extensive homology to EGF.

• Normally, HER-2 help to control how a healthy

breast cell grows, divides & repair itself.

• Mutation involving amplification of HER-2 convert c-

onc to onc.

• Over expression of HER-2 – Breast cells grows &

divide uncontrollably.
 4) H-TERT
• H-TERT gene code for TERT enzyme Telomerase Reverse Transcriptase.

• Telomerase maintain the telomere ie, the tandem repeats on DNA end.

• Telomere protect DNA from abnormally sticking together or breaking down.

• In each division, telomere no: shorten – After certain division telomere become so
short – Naked DNA ends signal for apoptosis.

• But in cancer, TERT gene is mutated – Its over expression leads to non stop supply
of telomere – So cell fails to do apoptosis.
 5) MYC GENE
• MYC gene code for CDK-Cyclin help in cell cycle,
apoptosis…

• This protein dimerise with related TF Max &

Regulate transcription of gene, that code for
products of cell cycle.

• Mutation due to Amplification – Over expression of

gene – CANCER.

• EG : Burkitt’s Lymphoma.
 TUMOUR SUPRESSOR GENE / ANTI - ONCOGENE

Proteins encoded by tumour

suppressor gene function in a diverse
array of cellular process, including :

• Controlling cell division

• Apoptosis

• DNA damage repair

• Study (in 1971 – Alfred knudson – in
Retinoblastoma) shows that chance
to develop the cancer follows a
dominant pattern of inheritance.

• It is due to heterozygosity for an

inherited loss-of-function mutation
in TS gene.

• A cancer develops only if a 2nd

mutation knocks out the function of
wild type allele of TS gene.
 1) RB GENE
• RB gene product ( RB protein / pRB ) is a 105 kilodalton nuclear protein, that is involved in
cell cycle regulation.

• Early in G1 phase, pRB bind to E2F TFs.

• So E2F cannot bind to specific enhancer on target sequence.

• So cell cycle factors encoded by these gene are not produced.

• DNA synthesis & cell division remains quiescent.

• Later in G1, pRB is phosphorylated through activation of CDK-Cyclin.

• pRB release E2F TFs.

• E2F bind to enhancer of target gene.

• Encodes proteins – So cell cycle progress through S-Phase & Mitosis.

• In cancer – RB gene is inactivated either by deletions or mutations.

• It abolish the ability of RB protein to bind with E2F TFs.

• So E2F always remain in active state – encode products – DNA synthesis & cell
division take place uncontrollably.
 2) TP53 GENE
• Code for 53-kilodalton TS protein p53.
• Li-Fraumeni Syndrome : Caused by inherited mutation in TP53.
• It is a rare dominant condition in which any of several different types of cancer may
develop.
• P53 have 3 different domains : Transcription Activation Domain (TAD), DNA Binding
Domain (DBD), Homo-Oligomerization Domain (OD).
• Mutations in DBD are typically recessive loss-of function mutation.
• If mutation is in OD – p53 molecules of mutant type dimerize with wild type p53 –
prevent its function as transcriptional activators.
• P53 is called as Guardian Of Genome – Associated with DNA Damage Repair &
Apoptosis.
 DNA DAMAGE
P53 & CELL CYCLE ARREST APOPTOTIC PATHWAY

P53 P53 TF induces

P53 TF induces
synthesis of p21 Synthesis of BAX

P21 BAX
P21 inhibit phospho-
Rylation of CDK

CDKs
Prb become in hypo-
Phosphorylated state
PRB HYPERPHOS-
pRB bind to & PHORYLATED
Inhibit E2F TF PRB
BAX antagonize BCL-2 (apo-
E2F BCL-2
E2F does not bind to Ptotic Repressor)
Taregt genes

No products for further TARGET GENE

Cell cycle
DNA CELL CYCLE
PROTEIN PRODUCTS APOPTOSIS PATHWAY PREVENTED
REPAIR ARRESTED
3) Papc / ADENOMATOUS POLYPOSIS COLI
• pAPC - 310 kilodalton protein - It control differentiation
& proliferation of intestinal epithelial cells.

• pAPC appears to regulate cell division, through its ability

to bind to β − catenin present in cell.

• β – catenin naturally bind to other proteins including TF -

stimulate genes which code for proteins for cell division.

• TF - catenin interaction is favoured when signals

impinging on cell surface cue the cell to divide.

• This signal is necessary bcoz the intestinal epithelium

lose enormous no: of cells / day ( 1011 ).
• Newly formed cells loose ability to divide as they move
away from generative part of epithelium to mature part
– so cells no longer receive signals for cell division.

• In the absence of these signals, pAPC forms a complex

with catenin & degrade it.

• It keeps catenin level low in mature cells – so catenin

no longer bind with TF to activate genes for cell division
– So cell proliferation is controlled.
• Mutation to pAPC cause Familial Adenomatous Poliposis (FAP) a cancerous
state.

• FAP mutation is inherited as autosomal dominant condition.

• In mutation, cell looses the ability to synthesize functional pAPC – So cells

loose the mechanism to control β – catenin levels.

• β – catenin level in cell increases – Promote uncontrolled cell division.

• Initially, Adenomas will be benign, but later, at least one of them become
malignant.
 4) pHMSH2
• pHMSH2 protein is the human homologue of DNA repair protein called mut.s
found in bacteria & yeast.

• It is associated with Hereditary Non-Polyposis Colorectal cancer (HNPCC), - A

dominant autosomal condition.

• They are characterized by the occurrence of a small no: of adenomas, one of

which eventually progress to cancerous condition.

• Cell lines that are deficient in mismatch repair develop a mutant phenotype,
that appears to drive the accumulation of mutations required for tumour
development.
 5) BRCA 1 & BRCA 2 GENES
• BRCA 1 was mapped to chromosome 17 in 1970 & is isolated
in 1994 – it encodefor 220 kilodalton polypeptide.
• BRCA 2 was mapped to chromosome 13 in 1994 & isolated
in 1995 – It encode for 384 kilodalton polypeptide.

• Each contain transcriptional activation domain – Protein interact physically with

other proteins, in particular with pRAD51, an eukaryotic homologue of RecA
(bacterial DNA repair protein).
• So pBRCAs are mainly involved in DNA repair in human cells.
BRCA1 has 22 exons spread over 100 kb & its 7.8-kb-long mRNA encodes a polypeptide of 1863
amino acids.

Researchers found 5 oncogenic mutations with the candidate coding sequence.

1) A 11-bp deletion in exon 2 that causes a reading frame shift.

2) A nonsense mutation in exon 11 that cause premature termination of polypeptide synthesis.

3) A 1-bp insertion in exon 20 that cause a reading frame shift.

4) A missense mutation in exon 21 that substitute arginine for methionine in protein.

5) A case in which Candidate gene transcript was absent, possibly because a regulatory mutation
had occurred in some noncoding element
• Chance to develop these cancers is inherited as a dominant allele, with high
percentage.

• This mutation account for 70 % of all cases of breast cancer & about 10 % of all
cases of ovarian cancer.

• It is found that, mice that are homozygous for knockout mutation in either of
BRCA genes die early during embryogenesis.

• Because many different inactivating mutations in BRCA genes are found in

human population, genetic counselling for families that are segregating these
mutations can be difficult.
 GENETIC PATHWAYS TO CANCER
• ONCOGENESIS is not the result of mutation in a single proto-oncogene or a single TS gene.
• Rather, tumour formation, growth, metastasis… usually depend on accumulation of mutations
in several different genes.
• Eg: Benign tumours of large intestine develop in individual having mutation in Apc gene –
progression of these tumours to cancers requires mutation in several other genes.

Inactivation Activation Inactivation Inactivation Inactivation

of APC TS of K-ras onc of TS on 18 q of TP53 TS of other TS

NORMAL DYSPLASTIC EARLY INTERMEDIATE LATE METASTATIC

INTESTINAL EPITHELIUM ADENOMA ADENOMA ADENOMA CARCINOMA COLORECTAL
EPITHELIUM CANCER
THANK YOU