Oncogenes & Tumour Suppressor Genes

Dr Usama ALAlami

Viruses, Cancer & Oncogenes DNA viruses (SV40) and RNA viruses (retroviruses) are capable of transforming cells they infect into cancerous ones.

Researchers discovered SRC gene in Rous sarcoma virus

Rous sarcoma virus causes myeloma in chickens

However, the SRC gene was not of viral origin (where did it come from?) The gene is brought under the control of the viral promoter 2 aberrant expression

The original cellular gene is the PROTO-ONDOGENE (Potential to cause tumour id abnormally activated)

Previous mechanism in humans = not evident Mechanism in humans = Insertional Mutagenesis

DNA copy of virus inserts into cellular genome near protooncogene abnormal activation of proto-oncogene tumoregenisis.

Viruses Associated With Human Cancers

Virus
DNA

Associated Tumour
Burkitts Lymphoma Nasopharyngeal carcinoma Cervical cancer

Epstein-Barr

Papilloma virus
RNA HIV-1 HTLV-2

Kaposis Sarcoma Hairy cell leukaemia

4

Cellular Oncogenes Mutated forms of a normal cellular proto-oncogene, they promote cell growth thus tumour formation

Chromosomal abnormalities associated with tumours are now known to involve oncogenes e.g. CML (t9,22) and Burkitts lymphoma (t8,14)

In both examples, the breakpoint for translocation coincided with the location of an oncogene (ABL on chromosome 9 and MYC on chromosome 8)
5

Double minute chromosomal abnormalities in breast cancer associated with MYC oncogene amplification 60-100 cellular proto-oncogenes identified Functions Of Cellular Proto-oncogenes Proto-oncogenes involved in basically all functions of growth, proliferation and differentiation Mutations in proto-oncogenes result in abnormal activation of cell growth in the ABSENCE of external stimuli (malignant transformation)

Functions Of Proto-oncogenes
External stimuli

Binding to cell surface receptor Signal transduction

Gene expression
7

[1]

Growth Factors

Growth factors exist as polypeptides or steroid hormones React with receptors stimulate or inhibit cell growth Enhance cell growth by: @ @ G0-G1 advance (EGF, PDGF, FGF) Progress through G1 (IGF)

In contrast, TGB- inhibits advance through G1 phase of the cell cycle

Abbreviations
EGF: Epidermal Growth Factor

PDGF: Platelet Derived Growth Factor

FGF: Fibroblast Growth Factor

IGF: Insulin-Like Growth Factor

Growth Factor Action Models

Growth Factor Action Models Cell 1 Growth factor
Stimulate cell 2

Cell 1 Growth factor

Stmulate cell 1

Paracrine secretions by cell 2 Stimulate secretion by cell1 PARACRINE

AUTOCRINE

10

[2]

Growth Factor Receptors

Second function for proto-oncogenes is encoding growth factor receptors (GFR) Link information from extracellular environment to different intracellular signalling pathways e.g. Steroid receptors, haemopoiesis growth factor receptors and transmembrane receptor tyrosine kinases When growth factors bind to transmembrane receptor kinase activate receptor transmit signal to the nucleus

11

[3]

Signal Transduction

Several proteins capable of transacting with tyrosine kinase Examples include ABL and SRC RAS = best studied signal transducer RAS = H, K and N RAS subtypes

RAS GTPase activity signal transduction

12

[4]

Transcription Factors

Act on DNA to control gene expression This is final site of action for messages sent sent from growth factors Also final level at which control of growth and proliferation ultimately operates e.g. FOS, JUN and MYC MYC = C, N and L MYC MYC has carboxyl terminal DNA-binding domain MYC dimerizes with MAX Binding to DNA occurs at the CACGTG sequence
13

This sequence is promoter (start point) site for many genes regulated by MYC
Overexpression of MAX inhibition of MYC/MAX dimerization activity Inhibition of transcriptional activity of MYC Aberrant expression of MYC is evident in many tumour types FOS/JUN heterdomerize increased expression following exposure to PDGF and EGF

14

Mechanics Of Oncogene Activation

Insertional mutagenesis (already mentioned) When MYC is translocated from chromosome 8 to 14 (site of Ig) in Burkitts lymphoma, constitutive expression of MYC results. This results in continuous cell proliferation No single mechanism for activation of oncogenes e.g. RAS activated by point mutations and amplifications

15

Mechanism Of Oncogene Activation

Method Of Activation
Amplification Insertional mutagenesis Translocation Structural alteration

Oncogene
MYC, ERBB2 INT1, INT2 MYC ABL, KRAS
16

Detection Of Mutation
@ @ @ RT-PCR Direct sequencing Southern blotting

17

Tumour Suppressor Genes (TSG)

Genes involved in control of abnormal cell proliferation Also known as ortho-genes, onco-suppressor genes and antioncogenes

The Retinoblastoma Model

Retinoblastoma = childhood cancer occurring in two forms + affects the retina

40% of cases are hereditary

Two hot hypothesis
18

The Retinoblastoma Model: Continued .

Inherit one mutation in the germ line (FIRST HIT) First hit = phenotypically harmless (SECOND HIT) = occurs in retinal cell = tumour develops Mutation analysis studies revealed deletion of the retinoblastoma gene (RB) on chromosome 13 (second hit) + assumed mutation in first copy on chromosome 13 (first hit) RB gene encodes RB protein with DNA-binding activity

Viral proteins bind to RB and inactivate it allow uncontrolled cell division to occur

19

Phosphorylation Of Retinoblastoma

RB Dephosphorylated Active Phosphorylated Inactive

20

Hereditary Cancers
{1} Wilms Tumour

Rare childhood renal tumour

Deletion of 11p13 Associated with genito-urinary abnormalities + mental retardation

11p13 is the site for WT1 gene

WT1 binds to early growth response gene 1 (EGR1) and suppresses it suppress growth TSG

IGF2 also implicated in Wilms tumour

21

{2}

Familial Adenomatous Polyposis

Polyps in colon + rectum mainly Turns malignant Translocated Adenomatous Polyposis Coli (APC) gene = TSG on chromosome 5q21

22

P53: The Guardian Of The Genome

p53 located on chromosome 17p Identified ten years ago because of involvement of chromosome 17 in a wide variety of cancers p53 functions as cell cycle checkpoint protein

It works alongside genes that have growth-suppressing activities

It functions in the G1 phase of the cell cycle

p53 also induces apoptosis (programmed cell death)

p53 prevents or repairs gene amplification p53 Interacts with DNA replication factor (RPA) and inhibits 23 its binding to ssDNA Inhibit DNA replication and growth

P53 as transcriptional Transactivator

p53 inhibits cell growth by promoting transcription of certain genes

The p53-activated genes suppressing activities

encode

proteins

with

growth

1993, a gene called WAF1 was shown to be induced by p53

WAF1 encodes p21 protein

Overexpression of p21 protein growth suppression

WAF1 identical to CIP1

24

Inactivation Of p53
Mouse double minute 2 (mdm2) cellular oncogene encodes p90 protein p90 interacts with amino terminal of p53 inactivation

Interaction Between Oncogenes And TSG In the Colorectal Cancer Model

Colorectal cancer has several hereditary forms Well-defined pattern of progression from adenomas to carcinomas

25

Interaction Between Oncogenes And TSG In the Colorectal Cancer Model. Continued
Genes involved are: a) APC

b)
c) d)

KRAS
DCC (Deleted in colon cancer) p53

26

Colorectal Cancer Model

Normal epithelium

5q mutation or loss APC gene

Hyperproliferative epithelium Early adenoma

12p mutation KRAS

Intermediate adenoma

18q loss DCC

Late adenoma

17p loss p53

Carcinoma

Other alterations
Metastases
27