Cancer Biology:

Tumorigenesis can stem form errors in signaling pathways

and cell cycle regulation mechanisms that we have discussed.

Objectives:
•identify the characteristics of cancer cells
•describe the hallmarks/acquired capabilities of cancer cells
•describe the multi-hit model of tumor development
•identify the categories of genetic targets
•differentiate between tumor suppressors and oncogenes
 Stages of tumor development
Benign tumor
–slow growth, localized
–closely resemble normal
cells
–express normal cell-specific
markers
normal epithelial cells
Malignant tumor
–divide rapidly
–high nucleus/cytoplasm ratio
–less differentiated than
normal cells
–loss of cell-specific markers
preinvasive cancer cells –invade surrounding tissue
Pap smear to detect cervical cancer:
Human Liver Tumor

High nucleus/cytoplasm ratio in dark

staining tumor cells
Lodish Figure 23-1
Metastasis
–Spread of tumor cells, enter circulatory system
–proliferate & establishment of 2º area of tumor growth
–Loss of cell-specific markers

Metastasized melanosarcoma invading the liver

 The Hallmarks of Cancer

D. Hanahan and R. Weinberg.

2000. Cell.100: 57-70

Which genes are disrupted, resulting in each of these

acquired characteristics? hallmarks
Genetic basis for cancer:
1st evidence that genetic material can cause tumorigenesis:
DNA from cancer cells transforms cultured cells:

Oncogenic
transformation

Transformed cells:
similar to tumor cells
Altered morphology,
reduced need for
growth factors,
altered chromosome
complement
(aneuploid), avoid
apoptosis
 Oncogenes

•Any gene that encodes a protein capable of transforming

cells in culture or inducing tumors in animals

•Normal cellular gene called a proto-oncogene

e.g.
proto-oncogene = ras, the normal cellular gene
oncogen = rasD , codes for constitutively active Ras protein

But, is one oncogene enough?

Two broad classes of genes:

proto-oncogenes: mutants (oncogenes)

= increased activity

tumor suppressors: mutants

= loss of restraint

Loss of cell regulation that gives rise to cancer is due

to genetic damage in somatic cells, but increased
susceptibility linked to inherited mutations
Multi-hit Model:
Evidence indicates that tumorigenesis is a multistep
process that reflect genetic changes

Evidence: 1) analysis of tumours

A tumor is derived from a single cell (clonal)

Analysis of DNA from cells within a human tumor shows

that tumor cells share the same genetic mutations
(somatic mutations).

multihit
Human Colorectal Cancer

First step - loss of function of APC (tumour suppressor),

benign polyps. See increased cell proliferation.

Second step - cells then get some or all of the other

mutations in varying order, RasD, DCC, p53.
See uncontrolled cell proliferation

Third step, Fourth step - mutations in tumor suppressors

and/or Oncogenes
Leads to malignant carcinoma and metastasis
Multi-hit Model:
•Evidence 2) epidemiological studies

Cancer incidence  exponentially with age, mutations

accumulate in multiple genes over time
Multi-hit Model:
Evidence: 3) mouse model Cooperativity of Oncogenes:

Myc or Ras
activation alone:
tumors occur in
mice at a low rate

High rate of
tumorigenesis
seen with more
than one oncogene

Long latency of tumor formation also reflects need to

acquire multiple mutations (not just in ras and myc)
Lodish Fig. 23-6
Parallel pathways to tumorigenesis:
The genetic basis of cancer:

Proto-Oncogenes & Tumor Suppressors

General:

Cancer cells acquire autonomy from external regulatory

signals: lower requirement for growth-stimulatory
signals and diminished sensitivity to inhibitor signals
 Gain-of-Function Mutations in proto-oncogenes.

•Most proto-oncogenes are essential and conserved

•Required for activating cell division
•Activation of proto-oncogene to oncogene involves gain-
of-function mutation

Classes of proto-oncogenes:

Growth factors
Hormone Receptors (pathways that induce cell division)
Signalling proteins e.g. Ras
Transcription factors e.g. Fos, Myc
Anti-Apoptosis proteins (keep cells alive)
Loss-of-function Mutations in tumour suppressors.

•Tumor suppressors usually inhibit cell proliferation

(brakes of the cell),
cell) so loss of function => cancer

•Both alleles must be inactivated or lost to promote

tumor development

Classes of tumor suppressors:

Hormone Receptors (pathways that inhibit cell division)

Pro-Apoptosis proteins (remove damaged cells)
Cell cycle regulators e.g. p16, Rb
and Checkpoint Control proteins, eg. p53
DNA repair enzymes
Oncogenic Mutations:
Activation of growth-promoting proteins

(1) Growth factors

• increased production of normal growth factors by

cancer cells contributes to unregulated growth

• Virus-encoded activators of growth factor receptors

Spleen focus-forming virus (SFFV)
= Retrovirus, induces erythroleukemia
– Binds and activates Epo receptor on erythroid
progenitors (glycoprotein, gp55 binds Epo
receptor in place of Epo)
Oncogenic Mutations
(2) Growth Factor Receptors

(a) Activated Growth

Factor Receptors:

Deletion that removes

ligand binding
domain …
RTK gain-of-function
mutations that
cause dimerization
in absence of a
ligand.

Fig. 23-14
Oncogenic Mutations
(3) Activated Signaling Proteins
Constitutively active
Ras
• proto-oncogene is Ras, G-protein in RTK signaling
pathway
• oncogene is RasD, constitutively active Ras

•Mutation Gly-12 ---reduced GTPase activity, always

bound to GTP, always Active

•Ras oncogene is seen in many human tumors - about

40% of human tumors contain Ras mutations
(bladder, colon, mammary, skin, lung, leukemias,
neuroblastomas, pancreatic)
Mutations in genes coding for proteins that are in
the RTK-Ras pathway

General rule: the mutation will promote

tumorigenesis if it promotes cell division for
example by promoting the GTP-bound form of Ras

Are they tumor-suppressors or oncogenes?

Think about the normal function of the protein - is it

to suppress division (tumor suppressor) or promote it
(proto-oncogene)?
or
Think about what what would happen if there was a
loss of function mutation or a gain of function
mutation.
 Complete this table:

Gene Protein function TS/Oncogene?

GRB adaptor protein: SOS binding Onco
SOS (GEF) guanine exchange factor Onco

GAP GTPase accelerating protein TS

GDI Guanine dissociation inhibitor TS

hormone receptor? Onco

Raf Ras target Onco

MapKinase Kinase cascade target Onco

 Oncogenic Mutations

(4) Transcription Factors

Stimulate transcription of genes that promote progression

through cell cycle = targets of Ras-MAPKinase
pathway, e.g. fos and jun early response proteins

–oncogenic forms expressed in virally infected cells:

(v-jun, v-fos)
–expressed under constitutive promoters; 3’ deletions
increase longevity of RNA and increase protein
production
Tumor suppressors:
Loss of cell-cycle controls

Cell Cycle Regulators

•Cell cycle precisely regulated and controlled

•Ensures coordination of DNA synthesis, cell-size

changes & cytokinesis

•Malfunction of any cyclin or regulatory protein needed

for Restriction Point (RP) regulation or Check Point (CP)

= passage through RP or CPs and uncontrolled cell

growth
Tumor Suppressors: Control G1- S transition

Cyclin/CDK inhibitors:

•Mitogen withdrawal prior to restriction point causes

accumulation CDK inhibitors
•p27, p16: inhibits cyclin D/CDK = G1 arrest

Loss of functional p27

or p16 bypasses arrest.
Which mouse carries a
LOF mutation in p16?
 Tumor suppressor: Rb
–Binds E2F and prevents E2F-mediated transcriptional
activation of genes needed for DNA synthesis
–Hence required for G1 arrest

Loss of functional Rb bypasses checkpoint.

X
No inhibition of E2F E2F active
- no restriction point and
uncontrolled cell division in
modified from Figure 21-30 the absence of a mitogen
Are there other tumor suppressors or oncogenes associated
with the Restriction Point ?
Think about:
CycD : proto-oncogene
E2F : proto-oncogene
P16 tumour suppressor
Tumor suppressor: p53

Loss of functional p53?

•p53 mutations abolish

its ability to bind DNA
& activate expression
of p21 and apoptosis
genes

Bypasses arrest.
X

p53 is mutated in > 50% human cancers

 p53 is a cell cycle inhibitor … is it a tumor suppressor?

p53 : acts dominantly = oncogenic mechanism i.e. only

one mutant allele is required to have on oncogenic
effect. Why?

•active p53 is a tetramer of 4 identical subunits

•mutation in only one allele will cause defective p53 complex

Nicholls et al, 2002.

•DNA tumor viruses produce a protein that binds to and
inhibits p53
–Human papillomavirus (eg. warts, cervical cancer)
and monkey SV40 Large T

•Human carcinogens also inhibit p53

–e.g. Cigarette smoke (benzo(a)pyrene)
–60% of lung cancers have inactive p53
… Benzo(a)pyrene induces mutations in p53 at
mutational hotspots

Treatment? Express p53

Caveat? Excess p53 in mice is associated with accelerated aging