Apoptosis and

Cancer
 What is Apoptosis?
• Apoptosis is “programmed cell death”.

• Apoptosis is a normal physiological process:

– Foetal development: finger and toe development requires loss of
tissue between.
– Removal of endometrium at menstruation.
– Formation of synapses in brain requires loss of surplus cells.

• Apoptosis is need to remove cells that pose a threat:

– Cytotoxic T lymphocytes kill viral infected cells by inducing
apoptosis.
– Effector cells of the immune system must be removed after the
response.
– Damaged cells must be removed if they cannot be repaired.
 Apoptosis
• Apoptosis from Greek, meaning “to fall
away from.”

• The release of apoptotic bodies from

cells is supposed to be analogous to
“leaves falling from trees.”

• Pronounced A-po-toe-sis
 Discovery of Apoptosis
• Kerr, J.F.R, Wyllie, A.H. Currie, A.R., 1972. Apoptosis:
a basic biological phenomenon with wide ranging
implications in tissue kinetics. British Journal Cancer
26, 239-257.

• They observed a “necrosis-like” process in the livers

of rats where the portal supply of blood had been
interrupted.

• No inflammation was observed (typical of necrosis).

• Histologically, the cells appeared as small particles

of chromatin surrounded by cytoplasm.
 Apoptosis in Cells
• Cells that undergo apoptosis:
– Shrink.
– Develop bubble-like blebs on their surface.
– Chromatin (DNA + protein) begins to degrade.
– Mitochondria break up releasing cytochrome c
– Cells break into small membrane wrapped fragment
– The phospholipid, phosphatidylserine is exposed on
surface
– Cell fragments are removed by phagocytosis.

• The process is very orderly and hence the name

“programmed cell death”

• An intrinsic property of normal cellular metabolism.

What Does it Look Like?
 Defects in Apoptosis
• Defects in the regulation of apoptosis
contributes to many diseases.

• Diseases where cell accumulation occurs

– Cancer

• Excessive cell loss

– Stroke
– Heart failure
– Neuro-degeneration
– Aids
 What Causes these
Changes?
• The changes in cell morphology are caused by the action of
proteases.

• These are cysteine proteases which cleave proteins at

aspartic acid residues. These are known as caspases.

• Cysteine Aspartate Specific ProteASEs

• These proteins are present as inactive zymogens in

essentially all cells.

• Proteolytic cleavage of the caspases at conserved aspartic

acid residues activates enzymes of 10 and 20kDa subunits.
 Mechanisms of Apoptosis
• Three different mechanisms by which a cell commits
suicide by apoptosis.

1. Generated by signals arising within the cell

(Intrinsic).
2. Triggered by death activators binding to receptors at
the cell surface (extrinsic):
– Tumour necrosis factor-α (TNFα )
– TNF-β (Lymphotoxin)
– A molecule that binds to the Fas cell surface receptor
(Fas ligand: FasL)
1. Triggered by cell damage by, for example, reactive
oxygen species (extrinsic).
 Intrinsic (Mitochondrial)
Apoptosis
• Outer membrane of mitochondria express the proto-
oncogene Bcl-2

• Bcl-2 is normally bound to a protein called Apaf1

– Apaf1: apoptotic protease activating factor 1

• Internal cell damage causes Bcl-2 to release Apaf1

which allows a related protein Bax to penetrate
mitochondria.
– Causes release of cytochrome c

• Cytochrome c, Apaf1 and caspase 9 form a complex

– Apoptosome
 Intrinsic (Mitochondrial)
Apoptosis
• Apoptosome complexes aggregate in the
cytoplasm

• Caspase 9 cleaves and activates other caspases

– Digestion of structural proteins in cells
– Degradation of chromosomal DNA

• Phagocytosis of cells.
 Pathway Summary

Apaf1
Bcl-2
Apoptosis

Mitochondria Bcl-2

Bax
Caspase cascade

Apaf1
Caspase 9
Cytochrome c
Internal Death Signal

Apoptosome
Extrinsic Mechanisms of apoptosis
(Death Receptor Pathway)
• The Fas and TNF receptors are integral cell
membrane proteins

• The binding of TNF or FasL (death activators) to the

cell membrane causes up-regulation and activation
of caspase 8.

• Caspase 8, like caspase 9 initiates a cascade of

caspase activation leading to phagocytosis of the
cell undergoing apoptosis.
Death Receptor Pathway
DNA damage and Apoptosis
 P53 and Apoptosis
• Bcl-2 and Bax genes regulate the release of
cytochrome c.

• The promoter for the Bax gene contains P53

binding sites.
– Upregulated in response to DNA damage.
– P53 can also repress Bcl-2

• Reactive oxygen species are powerful inducers of

apoptosis.

• Fas:FasL interaction causes the caspase cascade

leading to apoptosis.
– Fas expression is thought to be P53 dependent.
P53 summary
 Apoptosis and Cancer
• Loss of P53 activity in some 50% of all human tumours can
disrupt the normal apoptosis pathway.

• HPV virus and cervical cancer E6 oncoprotein can also

disrupt apoptosis by degradation of P53.

• Epstein Barr virus associated with some lymphomas

produces a Bcl-2 homolog and stimulates Bcl-2 production in
infected cells
– Bcl-2 is an anti-apoptotic protein.
– Translocation (t14:18)

• Some B-cell leukaemias express high levels of Bcl-2

– Oncogenic translocation of Bcl-2 gene to region for antibody
production
 Apoptosis and Cancer
• Melanomas avoid apoptosis by inhibiting the
expression of Apaf1.

• Some lung and colon cancer cells express a protein

that binds to FasL.
– Cytotoxic T cells cannot bind and induce apoptosis.

• There are examples of cancer cells expressing high

levels of FasL.
– Can kill cytotoxic T cells since T cells express their own
Fas)
 Summary
• Define apoptosis

• Discovery of apoptosis

• What happens to apoptotic cells

• Routes to apoptosis
– Intrinsic (Mitochondrial)
– Extrinsic (death signal)
• Cytotoxic T cells mediated

• Role of P53 in Apoptosis

• Apoptosis and cancer

– Mechanisms of avoiding apoptosis