Cellular apoptosis and

disease

Zhao Mingyao
BMC.ZZU

2006 ， 06
 Cell signal transduction

cell change

signal

Apoptosis
↑or↓or loss→disease
 Concept of apoptosis

• ——autonomic and ordered cell

death controlled by gene or called
as programmed cell death (PCD)

model: the nematode,

drosophila
and mouse
Flow Cytometer
nematode drosophila mouse

research model
 12000

10000
publications

8000

6000

4000 rapid up-shift

period
2000
silent period
0

1970 1975 1980 1985 1990 1995 2000 2005

Thirty years on apoptosis
Classical
Apoptosis
model

• Nematode : total 1090 cells in

young ,959 cells in adult, 131
cells lost ?
• Time : 1 h r
• Gene : 14 gene known
Section1 Instruction of apoptosis
1.differences between necrosis
and apoptosis
Strong Weak ~
stimulation
Active
Passive new protein
ATP(-) ATP↓

溶酶释出，
炎症反应
? Apoptotic bodies

Comparison between apoptosis & necrosis

DNA electrophoresis

necrosis apoptosis
+

famous nucleosomal ladder

2.the biological meanings of
apoptosis
 Holes are forming ?

Mitochondrio-nuclear AIF translocation in the interdigital

cells of the mouse embryo
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Apoptosis importance
①Maintain normal development and growth:remove
excess cell
② Maintain homeostasis : remove damaged, mutant
senile cell
③Active defense function: block replication of cells
infected by virus
3. apoptosis process (period)
 course of cellular apoptosis
1.Transduction of apoptosis signal
starting signal Ca2+ 、 cAMP 、 ceramide
2.activation of apoptosis gene
express enzymes and other substance
3.execution of cellular apoptosis
DNase and Caspases
4.removal of apoptotic cell
engulfed by near cell ( Mφor other cell )
4.the two major biomedical
changes in the courses of
apoptosis
 (1)Endogenous Dnase
activation and its effect
• A series cellular ST activation
• Execution of chromosome DNA
cleavage
 • Fragment of DNA ： classic ladder pattern ？

chromosome
Endonucleases first cleave
5min

300 kb pieces

50kb

90min
180~200bp or its
whole folds fragment
(2) Caspases activation and
its effect
• A group of protease ( 13 members )
cysteine-containing aspartate-spicific protease

caspases

note:Cysteine 半胱 aa
Aspartate 天冬 aa
 Caspases function
①Deactivating inhibitor of apoptosis: as
Bcl-2
②Hydrolyzing pro structure, cellular
decomposing, apoptotic body
③ Hydrolyzing the related active
proteins : make them to gain or lose
function
Section 2 Regulation of cell
apoptosis
1.Signal pathway and effectors
of apoptosis
 [Ca2+]i dyshomeostasis TNFa, Fas
DNA damage ROS

p53
bax
Adaptor
(FADD,TRADD)
Cyto C leakage
Bid
Bad
Bcl-XL-Apaf-1 Apaf- cyto c- dATP Caspase 8

Caspase 9

Caspase 3

Cell apoptosis Signal pathway of apoptosis

 2. regulators of apoptosis
• (1)the central role of mitochondria in apoptosis
[Ca2+]i dyshomeostasis
DNA damage ROS

Bax open
PTP
p53 bax VDAC
Bcl-XL
closing
Cyto C leakage
Bad
Bid
Apaf- cyto c- dATP

Caspase 9
mitochondria damage lead to
apoptosis

PTP opening

(permeability transition pore)

Inducing factor →Δ Ψm↓

PTP
permeability↑→apoptosis
starting factor （ Cyt.
C ， Apaf ， AIF ） released
Abbreviation note:
•PTP----permeability transition pore
•Apaf----apoptosis protease activated factor
•Cyt.C----cytochrome C
•AIF---- apoptosis inducement factor
 Deleterious network hypothesis of cellular
apoptosis
– – Bcl-2 –
Mt Δ Ψm↓ PTP opening Apaf+Cyt.C

+
–
Caspase
inhibitor AIF
Pro-Caspase 9 Caspase-9
+
Pro-Caspase3 Caspase-3
Inactivity +
DNase +
DNnase Ca2+ activated Protein hydrolyzed

DNA rupture apoptosis

 (2)bcl-2 family pro
• Bcl-2, Bcl-XL, Mcl-1 promote survival
(anti-apoptotic pro)
• Bax, Bad, Bak, Bid induce death
(pro-apoptotic pro)

cell
 bcl-2
• Binding Apaf-1
• Prevent release of cyto C
• Regulator of Ca2+ homeostasis
• antioxidant
• Inhibiting ~ ： bcl-2
• Promoting ~ ： fas ， P53
• Dual direction regulating ~ ： c-
myc ， bclx
Bcl-2 —— 229aa （ mouse 236aa ）
distributing in endomembrane surface
Bcl-2----B cell lymphoma/leukemia-2
 (3)IAP pro
• (IAP)/BIRP gene family: XIAP, c-IAP-1, c-IAP-2, ILP-2,
NAIP, Livin and Apollon
• block apoptosis
directly interacting with initiator and effector caspases
preventing their proteolytic processing and enzymatic
BIRC# for "baculoviral IAP repeat-containing".

activity

BIRC# for "baculoviral IAP repeat-containing".

 The "Inhibitor of Apoptosis Proteins"
family
Characterization of IAP family members
• BIRC1 (Neuronal apoptosis inhibitory protein; NAIP)
• BIRC2 (API1; HIAP2; cIAP1; MIHB)
• BIRC3 (API2; HIAP1; cIAP2; MIHC)
• BIRC4 (XIAP; API3; MIHA; ILP)
• BIRC5 (Survivin; API4; TLAP)
• BIRC6 (Apollon; BRUCE)
• BIRC7 (MLIAP; KIAP; Livin)
• BIRC8: (ILP-2; TIAP)
•
******Inhibitory factor ：
cytokine— IL-2 ， NGF
hormone— ACTH 、 testosterone ,
estrogen
other— Zn2+(zinc), phenobarbital ,
Cysteine protease inhibitor, EB
virus, neutroamino acid
 3.Inducers of apoptosis
• (1) apoptosis induced by death signal

TNFa, Fas

Adaptor
(FADD,TRADD)

Caspase 8
 apoptosis signal pathway
cell
Induction
signal
many
ceramide pathways SAPK/JNK
RR NF-kB,JNK/AP-1 pathway

ROS + bax or
P53 down regulating
gene bcl-2
mtDNA
(2) apoptosis induced by DNA
damage
DNA damage

p53 bax

Cyto C leakage
Bid
Bad
Bcl-XL-Apaf-1 Apaf- cyto c- dATP
 oxidative stress induce
apoptosis?

• P53 gene activation

• activating Ca2+ /Mg2+ -dependent Dnase
• Ca2+ influx↑
• activating NF-kB and Ap-1 （ Fas)
(3)Other apoptosis triggers
1)Inducing factor ：
hormone, GF↑↑, GC
physic and chemic factor
immune
microbe
2)P53

molecular policeman? Blocking IGF+R

+ Bax Bcl-2↓

Cyclin D

p53
G1
Cyclin B
M S
Cyclin A+CDK

Cyclin A+CDK1
G2
 3) [Ca2+ ] dyshomeostasis

• activating Ca2+ /Mg 2+ -dependent Dnase

• activating nuclear transcription factor related to
apoptosis
• Ca 2+ promoting to exposing enzymolytic sites
between nuclear ribosomes
Section 3 abnormal cell apoptosis
in diseases
 1.tumor

insufficiency of apoptosis
• Bcl-2 express↑
• P53 mutation or loss
 2. immune diseases
(1) Autoimmune diseases
• Hashilmoto’s thyroiditis (HT ) Fas/FasL
mediated apoptosis
• SLE
 (2)AIDS
HIV CD4+ cell damaged selectively
 mechanism
of CD4+ lymphocyte apoptosis
CD4+ Cell level

syncytia

CD4+
Effect and target effect

HIV
Apoptotic body
Signal substance AICD
HIV induce CD 4
+
apoptosis
Receptor,molecule level

gp120- R

Fas-R↑
TNF
Mφ
OFR

tat
Host cell CD4+cell
3. Heart failure
myocyte hypertrophy
and widening of
interstitial spaces
due to depostion of
collagen,
inflammatory cells
and amyloid.
 atherosclerosis, AS
Insufficiency and excess of apoptosis exist together

oxLDL ↑
Endothelia ~↑
activated plt ↑
Smooth ~ insufficiency
Ag II
(proliferation↑>apoptosis↑ ）
hypertension

Endothelia barrier↓
 4.Alzheimer disease, AD

Neuron retrogression
—— loss of neuron in hippocampus
and basal nuclei
• Loss of Cholinergic neurons reachs
30%~50% ， involved cortex
 AD mechanism
disease cause such as OFR
neuron

Ca2+ inward flowing↑

activating related genes

promoting β- amyloid content ↑

Tan pro overphosphorylation
Section 5 regulating apoptosis in
treating diseases
 meaning of apoptosis in prevention and
treatment of diseases

• (1) Correctively administration of the related factors

to ~
• Inhibiting ~ ： AD---- NGF
• acute T cell leukemia ： low dose
radiation 、 TNFα
promoting ~ : heat therapy of local tumor ；
high temperature 43℃ 、 30min
• prostate glands cancer ： removal of
testicle(testes), testosterone↓
 (2) Interfere apoptosis ST
• FAS/FASL ST system ： adriamycin
stimulation
• SPP （ metabolism products of
ceramide ST system ）： transducting
proliferation
signals ， antiapoptosis→AIDS ， AD
 (3) Regulating related gene to
apoptosis
• transfection of wild type P53 gene
 (4) Control related enzyme to
apoptosis
• DNase ： Ca 2+ /Mg 2+ -dependent
Ca 2+ ↑ activation ；
Zn 2+ ↑inhibiting
• Caspases: Ca2+ directive or indirective
activation
 (5)Prevent Mt Δ Ψm↓
• Cyclosporin A,immune inhibitor blocking Δ
Ψm↓ ， its depriver without immune
inhibiting effect, but stabilize Δ Ψm,
prevent↓
• A simple experiment
• A smart observation
• A further thinking
• Show a very different formation of
cellular death and it`s abstruse
meaning of life

review founding the process of cellular apoptosis

THE 2002 NOBEL PRIZE WINNER

Sydney Brenner H. Robert Horvit John E. Sulston

1/3 of the prize 1/3 of the prize United Kingdom
United Kingdom USA The Wellcome
Trust Sanger
The Molecular Massachusetts
Institute
Sciences Institute Institute of
Cambridge,
Berkeley, CA, USA Technology
United Kingdom
(MIT)
b. 1927
Cambridge, MA, b. 1942
(in Union of South
USA
Africa)
b. 1947
cell lineages of
nematode Apoptosis gene the nematode
Sydney Brenner receiving his Nobel Prize from His
Majesty the King at the Stockholm Concert Hall.
Photo: Hans Mehlin, Nobel e-Museum
THE NOBEL DIPlOMA
 support & cooperation in my teaching

more success
on the way of study & work in future

Zhao Mingyao(Jasper)

2006-06-08