THE CELL CYCLE

 It is the period from the time a cell

comes into existence until cell divides
into 2 daughter cells.
 It takes 20 to 24 hours in rapidly
growing cell cultures.
 In the human body it may be
 fast as in cell culture (hemopoietic cells)
 may take 6-12 months (liver cells).
 Phases of Cell Cycle
1. The gap 1 (G1) phase
 Cell growth occurs.
 It takes 9 hours in rapidly growing
cells.
 It may take several months(G0 phase).

2. The synthetic (S) phase

 DNA synthesis occurs.
 It takes 6-9 hours.
3. The gap 2 (G2 )phase
 The cell prepares itself for mitosis
 It takes 4 hours.
4. The mitotic (M) phase
 Cell division occurs
 It takes 1-2 hours.
 It includes nuclear division ( karyokinesis)
followed by cytoplasmic division (cytokinesis).

NB. The Gl, S, and G2 phases are called the interphase.

REGULATION OF THE CELL CYCLE

Cyclins (family of proteins) regulate the

transition of a cell from one phase to another.
The cyclins concentration increases and
decreases during different phases of the cell
cycle
The cyclins activate certain cyclin-
dependent protein kinases (CDKs) that
phosphorylate substrates essential for the
passage of the cell from one phase to another.
 Initiation of cell cycle
 It is initiated by the binding of a growth factor
to its receptor on the plasma membrane of the
cell.
 The growth factor receptor undergoes auto-
phosphorylation on tyrosine residues and
becomes active protein tyrosine kinase that
can catalyze phosphorylation of certain target
proteins on tyrosine residues.
 An intracellular signal finally induces the
production of cyclins.
 During the Gl phase: cyclin D increases late
in the Gl phase

 It forms a complex with CDK4 and

CDK6, called mitosis promoting factor
(MPF)
 MPF catalyzes phosphorylation of the
retinoblastoma (Rb) protein.
  The Rb protein is separated from transcription
factor E2F required for the transcription of
genes that code for proteins required to traverse
the Gl-S restriction point, (point beyond which
the cell continues to divide even in the absence of
the growth factor) which becomes active.

 MPF can be inhibited by cyclin kinase inhibitory

proteins (CIP) (p16 & p21) which are also known
as cyclin kinase inhibitor (CKI).

NB: The dephosphorylated Rb protein binds to and

inactivates E2F
  In early S phase: The E and A cyclins
activate CDK-2, which activates DNA
synthesis.
 During the S phase: cells contain large
quantities of enzymes required for DNA
synthesis.
 enzymes required for the synthesis of dNTPs
 thymidine kinase
 dihydrofolate reductase
 ribonucleotide reductase
 DNA polymerases
In G2 phase : B cyclins are produced late in the G2
phase.
*They activate CDK-1, a protein kinase
responsible for traversing the G2-M checkpoint.
*CDK-1 catalyzes the phosphorylation of:

(1) Histones: leading to condensation of chromosomes.

(2) Nuclear lamins: leading to disassembly of nuclear

membrane.
(3) Actin attachment proteins: leading to loss of
attachment and cell rounding up.
(4) Microtubules and cytoskeleton proteins: leading to
formation of mitotic spindle.
 Daughter cells should receive the same
genetic information possessed by the parent
cell to maintain genetic stability ( replication
must be complete and carried out with high
fidelity).
 Nuclear DNA is replicated only once during
the S phase .
 A pair of chromosomes replicates
simultaneously within a fixed period of the S
phase.
 DNA integrity is continuously monitored
throughout the cell cycle in 4 checkpoints.
 In G1 phase : Check DNA damage
 In G2 phase : Check DNA damage
 In S phase : Check completeness of replication
 In M phase : Check the proper chromosomal segregation

 If the damage could be repaired the cell cycle

continues, if it cannot be repaired the cell
undergoes apoptosis (programmed cell death).
 Apoptosis
 Apoptosis (dropping off) or cell suicide is a
programmed cell death that occurs during
 Embryogenesis
 Development
 Adult life.

 Necrotic cell death caused by cell injury due to

anoxia or radiation.
 Death receptors on the plasma membrane bind
death signals, instructing the cell to initiate
apoptosis.
 Death signals as tumor necrosis factor
(TNF) and Fas-ligand (CD 95 L) bind the
extracellular domain of the death receptor
and this allows binding of specific proteins
to the intracellular domain and promotes a
cascade of protein-protein interactions.

 This activates caspases 8 and 9, which then

activate other caspases.
 Caspases are proteases enzymes, they catalyze
the breakdown of cellular proteins.

 They activate a specific DNase called caspase-

activated DNase (CAD), which hydrolyzes the
linker DNA between the nucleosomes,
breaking the DNA into fragments.

 On electrophoresis, these fragments show a

characteristic ladder appearance.
 Apoptosis is also caused by intracellular
stress.
 Disruption of mitochondrial membrane
leads to the release of cytochrome c, which
along with apoptosis factor 1 (apaf-1),
activate caspase 9 and initiates the cell
death cascade.
 The tumor suppressor protein p53
(unstable protein) is a DNA-binding
transcription factor becomes stabilized by
DNA damage.
  In mild DNA damage, p53 induces the production
of the p21 that inhibits the action of all CDKs,
stopping the cell cycle and allowing for DNA
repair.
 If DNA damage cannot be repaired, p53 activates a
number of genes that induce apoptosis.
 Mutation in the p53 gene inhibits apoptosis and
predisposes to cancer.

 The bax protein shares in protein-protein

interaction ending in activation of apoptosis.
 The bax protein is inactivated by b-cell lymphoma-
2 (bcl-2) protein.
 bax is proapoptotic while bcl-2 is antiapoptotic
(oncogenic).
 Examples for apoptosis
 Menstruation: Estrogens and progesterone are the
main stimulators of endometrial growth. Withdrawal
of these hormones, due to atrophy of the corpus
luteum causes apoptosis and degeneration of the
endometrium and precipitates menstruation.

 Thymus gland atrophy: Glucocorticoids induce

apoptosis in thymocytes and causes atrophy of the
thymus gland in adult life. This action is mediated by
an intracellular glucocorticoid receptor.
 ONCOGENESIS
 Definition:

Conversion of a regulated cell into a

cancerous one with uncontrolled
growth and metastasis.
 Causes:
 mutations or underexpression of tumor
suppressor genes.
 mutations or overexpression of
protooncogenes.
TUMOR SUPPRESSOR GENES

 Products of these genes block abnormal

growth and malignant transformation.

 They are usually recessive; both copies of

the gene must undergo mutation to allow
for malignant transformation.
Examples of Tumor Supressor Genes
1. The p53, p21, p16, and bax Genes

 About 50% of human cancers have a mutated

p53 gene.
 Cells with a mutated gene fail to undergo
apoptosis upon damage of DNA.
 A mutation in the p16 gene known as
multiple tumor suppressor 1 (MTS 1) gene,
occurs in a wide variety of cancers.
2. Retinoblastoma (Rb) Gene
 Tissues express Rb gene:
 retina, osteoblasts and fibroblasts.

 Mutation of the gene may be inherited or acquired.

 Retinoblastoma is a rare childhood tumor of the
retina.
 The child inherits one mutated gene through the
germline and tumor results from acquired
mutation of the remaining normal gene.
3. WT1 and NF1 Genes
 WT1 gene
 Mutation causes Wilms tumor ( kidney tumor in
children).
 The protein product of this gene suppress the
transcription of certain growth factors involved in
the regulation of kidney development.
 NF1 gene
 One of the most commonly mutated genes in the
population.
 Mutation causes neurofibromatosis (1:3000
births ; 97% of cases are benign).
4. Genes of DNA Repair Enzymes

 Enzymes of DNA repair prevent tumor

formation by repairing DNA.

 Defective genes may cause tumors :

 Hereditary Nonpolyposis Colon Cancer (HNPCC)
 Xeroderma Pigmentosa
ONCOGENES & PROTOONCOGENES
 Oncogenes
 They are genes that cause cancer.
 Viruses causing cancer in animals

 Rous sarcoma virus which causes sarcoma in

chickens.
 It is a retrovirus with a DNA intermediate that
integrates into the host genome as a provirus.
 The viral DNA contains a gene called src (sarcoma
causing) that encodes a protein-tyrosine kinase
(PTK), which phosphorylates several proteins in
transformed cells.
 Some viruses are oncogenic in humans.
 These include DNA viruses
 Epstein-barr virus (EBV, may cause Burkitt's
lymphoma and nasopharyngeal carcinoma)
 human papilloma virus (HPV, may cause cervical
carcinoma)
 hepatitis B virus (HBV, may cause HCC)
 human herpes virus (HHV, may cause Kaposi sarcoma).
 They also include RNA viruses
 hepatitis C virus (HCV, may cause HCC)

 human T cell lymphotropic virus (HTLV, may

cause T-cell leukemia).
 Protooncogenes
 They are normal nonmalignant cells contain
DNA sequences homologous to viral oncogenes.
 They are active at some stage of development.
 Their product proteins are important for cell
growth and differentiation. They include:
 growth factors
 growth factor receptors
 signal transduction proteins
 transcription factors
 Causes of Conversion of a
protooncogene to an oncogene
1. Promoter or Enhancer Insertion
 Retroviruses insert their DNA into the host genome.
 If the viral DNA promoter or enhancer is inserted
near a protooncogene in the host cell it becomes a
hyperactive oncogene.
 An example is the insertion of a viral promoter or
enhancer near the inactive c-myc gene, leading to
avian leukemia.
2. Gene Amplification
 Increased number of copies of genes.

 Microscopically appear as homogeneously

staining regions (HSR) on the chromosome
or as extrachromosomal double minute
(dmin) chromosomes.

 A growth factor oncogene (hst) is amplified

in some cases of breast cancer.
 ErbB2 (HER2/neu) gene is amplified in 20-
30% of human breast cancer and is
associated with poor prognosis.
 N-myc is amplified in neuroblastoma and is
associated with poor prognosis.
 Dihydrofolate reductase gene is amplified in
cancer patients receiving methotrexate, an
inhibitor of the dihydrofolate reductase
enzyme.
 3. Point Mutation
 Ras protein (product of the ras protooncogene) is
found in all nucleated cells.
 It consists of the αs subunit of the G protein, which
has Intrinsic GTPase activity.
 Point mutation converts this gene into ras oncogene

with reduced GTPase activity.

 The effect of growth factors acting through G protein

continues after the growth factor dissociates from the

receptor.
 This mutation is observed in about 15% of cancers.
4. Chromosomal Translocation
 Reciprocal translocation between chromosome 9 and
chromosome 22 produces a fusion BCR-abl gene that encodes
a protein with high PTK activity.

 The new chromosome 22 is known as the Philadelphia

chromosome.
Thank you