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细胞凋亡. 细胞凋亡 Cellular Apoptosis 一、概论. 随着生物学研究的深入,生物学家越来越意识到细胞死亡,特别是细胞凋亡具有特殊的生物学意义,对于一个多细胞生物来说,要维持完整性和保持平衡性,凋亡是一个非常重要的生物学过程。多细胞生物的诞生、生长、发育、存活以及死亡,无一不伴随着细胞凋亡过程。.

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细胞凋亡

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Cellular Apoptosis


(embryonic development)Normal tissue turnoverselection of appropriate clones


1972KeerApoptosis

80

2002 H E



dying process(elimination process)

1

2squeeze

bleb

shrinkage


3

SEN

4Apoptotic bodies


5

chromatin

condenation

margination

compaction

fold


HE1000


HE1000



6

phagosome

lysosomal residual bodies


1<50%


2.Fesustrangluamirase


3


thrombospoudinFN


4,4-935%


the dying processthe elimination process


5

tingible body macrophages

civattebodies;

councilman bodies


civattebodies


H.E 10 50


apoptotic body

H.E 10 40


receptor-mediated cellular signalling pathways

180-200bpDNA


1endonucleaseDNAinternucleosomal linkage region

(Nick),DNADNADNA180-200bpDNA

2tissue tranglutaninase TTG)lr-TTGinvolucrin)


3calcium-dependent proteinase)

spontaneousteratogenXenobiotics


(caspases)

caspase

death receptorDR

caspase


1

death ligandsDRTNFFasLTRAILApo3LTNF-RFasCD40OX404-1BB80(death domainDD)


Fas/FasL

FasLFasFasFADDcaspase-8caspaseDDFADDFas LFas


TNF

TNF157T TNF-R1 TNF

TNFTNF-R1TNF - R1


2

(adenine nucleotide transporterANT)(voltage-dependent anion channelVDAC)bcl-215KD1.5KD


3 :

;

;


caspase

cCyt-ccaspase)

caspases/IAP(Smac/Diablo caspase )

(apoptosis inducing factorAIFDNA)

(inhibitor of apoptosis proteinsIAPcaspase)

-G(endonuclease G)


c Cyt-c

Cyt-c

Cyt-c1(apoptotic protease activating factor 1Apaf-1ced-4) caspase-9caspase-3caspases


( apoptosome)

(death complexes)Cyt-cSmac/DiabloCyt-cApaf-1(heptameric apoptosome)caspase(caspase recruitment domainCARD)caspase-9caspase-3caspase-6caspase-7




2Cytokines:

5

1growth factors

2colony stimulating factors

3 interleukin

4tumor necrosis fctors TNF

5 interferon





TB

95%TCTCRTCRTCR


BTCSIGBCBC

WBCWBCWBC



1

2

3 >,


Apoptotic index AI

basel cell carcinoma

ER/PR+,BCl-2


prostatic specific antigen(Androgen drpendent prostatic carcinoma ADPC )BCl-2 Androgen Independent prostatic carcinoma AIPC BCl-2+70%3AIPCsuraminAIPC


10-25%BCl-2BCl-2

BCl-2 BCl-XLBCl-XLBCl-I


[p35p53 RBDOCWT-1 1 wimls tumor type-1DAD 1]

[suicidal gene

]

APO-1/FasTGF-SP2Bax c-relBel-XSBak

2 c-myc c-eld 1ras v-src

/

Bcl-2EIB LMW-5-HLC-Kit Bel-XL


1

2

3


BCL2Caspase IAP Fas P53Cmyc


1.Bcl-2

Bcl-2Bcl-24(BH1-4)


1 Bcl-2


BaxBcl-2Bcl-2BaxBcl-2Bax/


4 Bcl-2

Bcl-2Bcl-2Bcl-2Bcl-2l0080TB6540804080752510--15


85t(1418)(q32q21)1418bcl2bcl2BBcl-2

Bcl-2MALT


Bcl-2


Follicular lymphoma (Bcl-2 immunostaining)


Follicular lymphoma: Bcl-2



Bcl-2


Bcl-2 : ?


CD3 T


2. Caspase

Caspase

Caspase2

.


Caspase

caspase-4(TX/ICH-2/ICEref-)

caspase-5(TY/ICEref-)

caspase-13(ERICE)

mcaspase-11(ICH-3)

mcaspase-12

caspase-1(ICE)

caspase-14(MICE)

caspase-3(CPP32/apopain/Yama)

caspase-7(Mch3/ICE-LAP3/CMH-1)

caspase-6(Mch2)

caspase-8(MACH/FLICE/Mch5)

caspase-10(Mch4)

caspase-2(ICH-1/mNedd2)

caspase-9(ICE-LAP6/Mch6)

CED3


Caspase

14Caspase3

1Caspase-2-8-9Caspase

2Caspase-3-6-7Caspase

Caspase

3Caspase-1-4-5


(2)Caspase

Caspase


3. IAP

IAP6IAP(NAIPc-IAP1hIAP-2c-IAP2hIAP-1 XIAP/hICPSurvivinBRUCE)TNFRFasVP16IAPcaspaseXIAP>c-IAP2c-IAPlSurvivin


5IAP

XIAP

clAPlcIAP2

NAIPRTPCR

SurvivinSurvivin


4Fas

APO-1CD951989NTFRNGFRN45000CD95FaslFas APO-1FaslTFasligantFaslFas


5P53

P538089P53P53

P53

G1


6c-myc

c-myc(HL-60)c-mycc-myc


7

ced3ced4. ICE(interleukin-l p-converting enzyme)ced3ICEBCL2

ced1ced2ced6ced7ced8

Nmc-1


()


Apoptosis Index(proliferatory Index)AI/PI


1P53WT

2P53, BCl-2

3

4

5

6


1

ab c

AIDS


2

aSLE bGN

ab


1972keer

1980DNA

1986caeovhabditis elegan1090131ced3 ced4


1

2

3


LM

PCLM

TEM

CLSM

SEM

FCM

TLP


2

Hoechs-PIHoechsDNAPI


3

DNA

DNADNADNADNADNA180-200bpDNADNA


4(ELISA )

ELISA DNA 5102/m L

:

(1);

(2) ;

(3)


5DNA

1993WifsmanDNA Fehsel1994TUNEL

:

(1)DNA TUNEL

(2)TUNEL


Tunel DNA-DNA


DAB

75 85 95

Tunel


TunelDAB125


TunelBCIP/NBT 250


RNA(RNase)

DNARNA DNARNA mRNA cDNA RNA

32PRNARNA RNA RNA mRNA


RT-PCR mRNA

RNA RNA(RNA sin) dNTPcDNA cDNAPCR PCR mRNA




1

2

3

4

5


40


1

Switch point gene


2Killer gene?

3

4

5 survival gene?


NN



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