BI 341 CHAPTER 13

MOLECULAR GENETICS OF CELL

CYCLE AND CANCER
Kim H. Brown, PhD
 Objectives
1. The Cell Cycle
2. Elements of cell cycle control:
a. Cyclins and CDK
b. Retinoblastoma Protein and EF2
c. Anaphase Promoting Complex (APC/C)
d. Checkpoints: p53, Mdm2, p21, bax
3. Cancer
a. Molecular factors: Proto-oncogenes and Tumor
Suppressors
b. Familial cancers: Li–Fraumeni Syndrome & p53,
Retinoblastoma, Leukemia

Cancer overview (long): http://www.cancercenter.com/what-is-cancer/

 Introduction to cancer
Leading cause of death in
Western countries.
• More than 1 million cases are
diagnosed US per year, and
more than 500,000 deaths.
Cancer is a genetic disease at
the somatic level
• characterized by gene
products derived from
mutated or abnormally
expressed genes.
• Some are inherited, but most
are created within somatic
cells that divide and form
tumors.
 Introduction Karyotype of a normal vs.
cancerous cell

Cancer usually arises from

mutations in many genes:
1. Single-nucleotide substitutions
2. large-scale chromosome
rearrangements

Affects multiple cellular functions:

repair of damaged DNA
1. Cell division
2. Apoptosis
3. Cellular differentiation
4. Migratory behavior CRISPR/CAS9 system as a
potential therapeutic:
5. Cell–cell contact https://www.youtube.com/watch?v=
O0usd9IZVCg
 Role of The Cell Cycle
1. To ensure that each chromosomal DNA molecule is
replicated only once per cycle
2. To ensure that the identical replicas of each
chromosome are distributed equally to the two
daughter cells
 The Cell Cycle
1. Under genetic control
2. It is a true cycle, it is not
reversible
3. Many genes are transcribed
during the cell cycle just
before their products are
needed
4. Mutations affecting the cell
cycle have helped to
identify the key regulatory
pathways
 The Cell Cycle
There are two major parts in the cell cycle:
1. Mitosis: M
2. Interphase: G1 = gap 1, S = DNA synthesis, G2 = gap
2
 Cyclins and cyclin-dependent protein kinases
(CDK): control progression of the cell cycle
• mitotic cyclins E, A, and B: periodic expression
• cyclin D is expressed throughout the cell cycle in response to
mitosis stimulating drugs (mitogens)
The cyclin-CDK complexes phosphorylate targeted proteins,
dramatically changing their activity

Cyclins and
CDKs:
http://highered.m
heducation.com/
sites/983409233
9/student_view0/
chapter10/stimul
ation_of_cell_rep
lication.html
The temporal expression pattern of activities of the
cyclin–CDKs in mammalian cells.
 Retinoblastoma (RB) controls the
initiation of DNA synthesis
RB halts cells at
G1 restriction
point by binding
(and inactivating)
the transcription
factor E2F
 If the cycling cell is growing
properly, cyclin-CDK complexes
inactivate RB by phosphorylation

http://highered.mheducation.com/sites/9834092339/student_view0/chapter10/h
ow_tumor_suppressor_genes_block_cell_division.html
Retinoblastoma: Insights and solutions
 Retinoblastoma: Insights and solutions

1. Replication error during mitosis

2. Chromosomal nondisjunction during mitosis (monsomy # 13)
3. Chromosomal deletion of RB1 gene
Retinoblastoma: Insights and solutions
Retinoblastoma: Insights and solutions
The progression from G2 to M is controlled by
a Cyclin B-Cdc2 complex known as
maturation-promoting complex

Note Cdc2 ~ Cdk1

Anaphase Promoting Complex (APC/C): adds ubiquitin
to its target proteins and marking them for proteolysis,
eliminates proteins used in the preceding phase as well as
proteins that would inhibit progression into the next
 Checkpoints
Checkpoints: maintain the correct order of steps causing
the cell cycle to pause while defects are corrected or repaired
 Three Main types of Checkpoints
1. A DNA damage checkpoint
2. A centrosome duplication checkpoint
3. A spindle checkpoint
 A DNA Damage Checkpoint
A DNA damage checkpoint arrests the cell cycle when
DNA is damaged or replication is not completed. Acts at:
1. G1/S transition
2. S period
3. G2/M boundary
 A DNA Damage Checkpoint
p53 transcription factor: activated by DNA damage

Protein Mdm2 keeps p53 inactivated in undamaged cells

by preventing phosphorylation and acetylation of p53 and
by exporting p53 from the nucleus
 A DNA
Damage
Checkpoint
Activated p53 triggers
transcription of a
number of genes –
p21, 14-3-3σ, Bax,
Apaf1, Maspin,
GADD45
DNA damage blocks cell cycle
progression
 A DNA Damage Checkpoint
DNA damage also
triggers apoptosis
(programmed cell
death):

In apotosis, a cascade of
proteases called
caspases are initiated
that culminates in cell
suicide
Centrosome
Duplication &
Spindle
Checkpoint
Monitors assembly of
the spindle and its
attachment to
kinetochores during
mitosis in the interest
of maintaining normal
complement of
chromosomes
 Cancer: some facts
1. Cancer cells have a small
number of mutations that
prevent normal checkpoint
function
2. Cancer is not one disease
but rather many diseases
with similar cellular
attributes
3. All cancer cells show
uncontrolled growth as a
result of mutations in a
relatively small number of
genes
4. Cancer is a disease of
somatic cells Capabilities acquired by cancer
cells
 Cancer

• 1% of cancer cases are familial:

show evidence for segregation of a
gene in pedigree
• 99% are sporadic: the result of
genetic changes in somatic cells
• Within an organism, tumor cells are
clonal, which means that they are
descendants from a single ancestral
cell that became cancerous
 Cancer Cells vs. Normal Cells
Cancer cells have lost
contact inhibition
(prevention of growth
and division by cell
contact)

Cancer cells have high

levels of telomerase,
which help to protect
them from senescence
(normal decline in cell
division), making them
immortal
Animation: http://highered.mcgraw-
hill.com/sites/9834092339/student_view0/chapter14/telomerase_function.html
 Telomeres and the cells life span (ch 3)
• Shortening of telomeres (repeat sequences at the ends of
chromosomes) owning to ineffective lagging strand
synthesis at the chromosomes ends limits cell cycle to 50-
70 divisions. Telomerase activity can elongate this life
span.
 Key Mutational Targets
Many cancers result from
abnormal control of the G1-to-
S transition
• These mutations often affect
apoptosis through their
interactions with p53
Mutational targets
• Proto-oncogenes which
promote cell division or
prevent apoptosis
• Tumor-suppressor genes
which prevent cell division
or promote apoptosis
Proto oncogenes will be more highly expressed in cancer cells (making
these type of mutations oncogenenic). Tumor suppressor gene
expression will be reduced (relative to wild type) in cancer cells.
 Oncogenes
Oncogenes (derived from proto-oncogenes) are gain-of-
function mutations, meaning the mutation increases
expression of genes that promote cell proliferation or inhibit
apoptosis
• Mdm2: overexpression leads to inactivation of p53 gene.
Mutants found in many tumors of adipose tissue, soft tissue
sarcomas, osteosarcoma, and esophageal carcinoma
• Cyclin D and CDK4: overexpression leads to unscheduled
entry to S phase. Mutants found in many esophageal
carcinomas, bladder and breast cancers
 Oncogenes: Growth factor
receptors
Growth factors bind to their receptor at the cell
membrane and activate Ras, cyclin D, and its partner
CDKs.
Epidermal Growth Factor Receptor (EGFR) mutants
have been found in brain tumors, breast and ovarian
cancers, head and neck cancers, and melanomas.
 Oncogenes: summary

Note: amplification can occur via mutation in promoter,

transcription factor, etc.
 Tumor-Suppressor Genes
Tumor-suppressor genes normally negatively control cell
proliferation or activate the apoptotic pathway

Loss-of-function mutations in tumor-suppressor genes

contribute to cancer progression.
 Tumor-Suppressor Genes
Loss of function of p53 Mutant p53 proteins are
eliminates the DNA checkpoint found frequently in
• Damaged proliferate, melanomas, lung cancers,
genetic instability increases colorectal tumors, bladder
the probability of additional and prostate cancers.
genetic changes,
• p53 is nonfunctional in more
than half of all cancers

Animation: The Tumor Suppressor Gene,

TP53.htm
 Tumor-Suppressor Genes

• p21: Loss of function

results in DNA
synthesis without
mitosis, increasing
ploidy
• Occurs in some
prostate cancers.
 Tumor-Suppressor Genes
RB: Loss of RB function frees E2F, hence, excessive rounds of
DNA synthesis are continuously being initiated.
 Loss of RB function is found in melanomas, small-cell lung
carcinoma, osteosarcoma, and liposarcomas
 Tumor-Suppressor Genes
Loss of Bax function prevents apoptosis. These mutations are
found in adenocarcinomas and in colorectal carcinomas
 Familial Cancers
Mutations that predispose to cancer can be inherited through
the germ line
• reduces the number of additional somatic mutations
necessary for a precancerous cell to progress to malignancy
 Li–Fraumeni Syndrome: p53
• autosomal dominant inheritance
• range of tumors, often more than one

A patient’s story: http://www.youtube.com/watch?v=knQM9JqU5BY

Dominant mutation of p53
 Retinoblastoma
RB is an inherited cancer syndrome which manifests
as a retinal tumor.
RB is caused by loss of heterozygosity in the tumor
cells, an event that requires 2 mutations
 Acute Leukemias
• Associated with uncontrolled proliferation of leukocytes.
• NOT consequence of alterations in cell cycle regulation or
checkpoints, nor are they familial
 promoter fusion: the coding region transcription factor is
translocated near highly productive enhancer
 gene fusion: The translocation breakpoints occur in introns
of genes for transcription factors in two different
chromosomes.
Transmissible
Cancers