Multi-Step Tumorigenesis

http://profhorn.aos.wisc.edu/wxwise/kti/bacteria/h5/sp3.html
 Tumor progression

Normal cells evolve into cells with increasingly neoplastic

phenotypes through a process termed tumor progression.
Tumor progression is driven by a sequence of randomly occurring
mutations and epigenetic alterations of DNA that affect the genes
controlling cell proliferation, survival, and other traits associated with
the malignant cell phenotype.
Multi-step tumorigenesis in a variety of organs
 Molecular alterations during human
colon tumor progression

* tumors bearing K-ras oncogenes rarely have

The precise contribution of mutant p53 alleles, and vice versa.
hypomethylation to tumor
progression remains unclear;
some evidence suggests that it
creates chromosomal instability.
Adenomatous Polyposis Coli (APC) is a regulator of several fundamental cellular processes, including cell
division, cell attachment, cell migration, cell polarization, and chromosome segregation during division.

One of the most recognized functions of APC is in regulating levels of beta-catenin, which is part of the WNT
signal pathway in cells.

T-cell factor

http://targetedcancercare.massgeneral.org/My-Trial-Guide/Diseases/Colorectal-Cancer/APC.aspx
 Alternative genetic paths during
colon cancer progression
Deleted in Pancreatic Cancer, locus 4

In all, 50% of pancreatic cancers, as

well as lesser proportions of other
cancers (such as bladder, breast,
lung, bile duct, and colon tumors),
experience a total inactivation of the
DPC4 gene due to mutations.
almost always the
1st inactivation in
the progression
Individuals with familial adenomatous
polyposis (FAP) start here !
Darwinian evolution and clonal successions
 Paradox
- A mutated, activated H-ras oncogene from a human
bladder carcinoma could fully transform NIH3T3 mouse
fibroblasts.
→ the acquisition of a single ras oncogene (a point
mutation on ras) sufficed to convert NIH3T3 cells
to a transformed, tumorigenic state.

- Why is cancer formation in humans a multi-step process ?

11.9 Multiple lines of evidence reveal that
normal cells are resistant to transfor-
mation by a single mutated gene

- The NIH3T3 cells are not truly normal. They constitute a cell
line, a population of cells that have been adapted to grow in
culture and to proliferate in an immortalized fashion.
- The results obtained from primary rat and hamster primary cells
(transfection of ras oncogene) were very different from those
observed previously with NIH3T3 cells. These primary cells
were not susceptible to ras-induced transformation.

- Single mutations are not sufficient for the development of cancers.

 11.10 Transformation usually requires collabor-
ration between two or more mutant genes
- A line of human promyelocytic leukemia cells carry both an
activated N-ras and an activated myc oncogene.
- When a myc oncogene was introduced together with an H-ras
oncogene into rat embryo fibroblasts, the cells become
transformed and tumorigenic.

Figure 11.23 The Biology of Cancer (© Garland Science 2007)

The ras and myc oncogenes have complementary
effects on cell phenotype

ras: anchorage independence

round and refractile appearance
loss of contact inhibition
(cytoplasmic mitogenic signaling cascade)
myc: immortalization
reduction of dependence on growth factors
(perturb cell cycle control machinery)
11.12 Human cells are constructed to be highly resistant
to immortalization and transformation

- Human cells rarely, if ever, become immortalized following

extended serial passaging in culture.
- The introduction of paired oncogenes (such as ras + myc)
could transform primary rodent cells, however, such pairs
consistently fail to yield tumorigenic human cells.
a possible explanation:
- The cells of laboratory mice carry extremely long telomeric
DNA (up to ~ 40 kb) and express telomerase activity.
- In contrast, normal human cells have far shorter telomeres
(5~10 kb), and most human cell types lack telomerase activity.
Human cells
+ hTERT gene (prevent telomere erosion)
+ SV40 large T gene (inactivate both pRb and p53)
immortalization

+ ras
morphological transformation in culture
(still unable to form tumors in nude mice)

+ SV40 small T gene (perturb the function of protein

phosphatase 2A, or PP2A)
tumorigenic

PP2A: The wolf in sheep’s clothing?

•June 2015
•Cancers 7(2):648-669
 Five distinct intracellular pathways involved in
human cell transformation

1. Mitogenic signaling pathway controlled by Ras

alterations are
2. Cell cycle checkpoint controlled by pRB commonly seen
3. Alarm pathway controlled by p53 in human
cancers
4. Telomere maintenance pathway controlled by hTERT
5. Signaling pathway controlled by protein phosphatase 2A
11.13 Non-mutagenic agents, including those
favoring cell proliferation, make impor
tant contributions to tumorigenesis

Are all mutagens carcinogens?

Are all carcinogens mutagens?

Indication of the importance of nonmutagenic

(nongenotoxic) carcinogens came from the study
of “tumor promotion” in 1940s.
 Induction of skin cancers requires certain
combinations of initiators and promoters

carcinogens, e.g.,
benzo[a]pyrene (BP),
7,12-dimethylbenz[a]anthracene (DMBA),
3-methylcholanthrene (3-MC)

TPA – 12-O-tetradecanoylphorbol-13-acetate,
a skin irritant prepared from the
seeds of croton oil

also called PMA

(phorbol-12-myristate-13-acetate)
from (E): The effects of promoter seem
to be reversible, thus, it may
exert a nongenetic effect on
the papilloma cells.
from (F): Additional treatments of the
promoter seem to push the
papilloma into a promoter-
independent state.
Explanation of the combined effect of
initiation and promotion
drive the proliferation
of ras* cells

The skin tumors that

emerge invariably
bear point-mutated
H-ras oncogenes,
indicating that this
mutation confers a
strong selective
advantage on cells in
the skin.
Why does TPA (a tumor promoter) promote
tumor formation?

TPA acts as a functional mimic

of diacylglycerol (DAG), which
activates protein kinase C.
DAG activates protein kinase C α (PKCα )

Figure 11.31 The Biology of Cancer

(© Garland Science 2007)
 Induction of inflammation by TPA

PKCα transgenic mice

wild type (keratin-5-promoter driven)

TPA was initially

chosen because it is
an irritant of mouse
skin and thus an
inducer of localized
neutrophil
inflammation.
infiltration
11.14 - 17 Toxic agents, mitogenic agents and
chronic inflammation can act as
tumor promoters
cytotoxic agents : damage cells → causing the proliferation of
the cells that have survived
the toxic effects
mitogenic agents : i.e.,steroid hormones
Estrogen and progesterone are involved in the
programming the proliferation of cells in
reproductive tissues
chronic inflammation – tissue damage from inflammation
– long-term anti-inflammatory drug use
links to reduced cancer incidence