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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


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(embryonic development)Normal tissue turnoverselection of appropriate clones


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1972KeerApoptosis

80

2002 H E


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dying process(elimination process)

1

2squeeze

bleb

shrinkage


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3

SEN

4Apoptotic bodies


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5

chromatin

condenation

margination

compaction

fold


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HE1000


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HE1000


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6

phagosome

lysosomal residual bodies


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1<50%


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2.Fesustrangluamirase


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3


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thrombospoudinFN


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4,4-935%


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the dying processthe elimination process


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5

tingible body macrophages

civattebodies;

councilman bodies


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civattebodies


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H.E 10 50


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apoptotic body

H.E 10 40


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receptor-mediated cellular signalling pathways

180-200bpDNA


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1endonucleaseDNAinternucleosomal linkage region

(Nick),DNADNADNA180-200bpDNA

2tissue tranglutaninase TTG)lr-TTGinvolucrin)


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3calcium-dependent proteinase)

spontaneousteratogenXenobiotics


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(caspases)

caspase

death receptorDR

caspase


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1

death ligandsDRTNFFasLTRAILApo3LTNF-RFasCD40OX404-1BB80(death domainDD)


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Fas/FasL

FasLFasFasFADDcaspase-8caspaseDDFADDFas LFas


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TNF

TNF157T TNF-R1 TNF

TNFTNF-R1TNF - R1


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2

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


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3 :

;

;


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caspase

cCyt-ccaspase)

caspases/IAP(Smac/Diablo caspase )

(apoptosis inducing factorAIFDNA)

(inhibitor of apoptosis proteinsIAPcaspase)

-G(endonuclease G)


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c Cyt-c

Cyt-c

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


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( apoptosome)

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


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2Cytokines:

5

1growth factors

2colony stimulating factors

3 interleukin

4tumor necrosis fctors TNF

5 interferon


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TB

95%TCTCRTCRTCR


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BTCSIGBCBC

WBCWBCWBC


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1

2

3 >,


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Apoptotic index AI

basel cell carcinoma

ER/PR+,BCl-2


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


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10-25%BCl-2BCl-2

BCl-2 BCl-XLBCl-XLBCl-I


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[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


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1

2

3


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BCL2Caspase IAP Fas P53Cmyc


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1.Bcl-2

Bcl-2Bcl-24(BH1-4)


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1 Bcl-2


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BaxBcl-2Bcl-2BaxBcl-2Bax/


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4 Bcl-2

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


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

Bcl-2MALT


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Bcl-2


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Follicular lymphoma (Bcl-2 immunostaining)


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Follicular lymphoma: Bcl-2


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Bcl-2


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Bcl-2 : ?


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CD3 T


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2. Caspase

Caspase

Caspase2

.


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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


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Caspase

14Caspase3

1Caspase-2-8-9Caspase

2Caspase-3-6-7Caspase

Caspase

3Caspase-1-4-5


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(2)Caspase

Caspase


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3. IAP

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


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5IAP

XIAP

clAPlcIAP2

NAIPRTPCR

SurvivinSurvivin


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4Fas

APO-1CD951989NTFRNGFRN45000CD95FaslFas APO-1FaslTFasligantFaslFas


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5P53

P538089P53P53

P53

G1


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6c-myc

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


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7

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

ced1ced2ced6ced7ced8

Nmc-1


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()


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Apoptosis Index(proliferatory Index)AI/PI


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1P53WT

2P53, BCl-2

3

4

5

6


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1

ab c

AIDS


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2

aSLE bGN

ab


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1972keer

1980DNA

1986caeovhabditis elegan1090131ced3 ced4


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1

2

3


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LM

PCLM

TEM

CLSM

SEM

FCM

TLP


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2

Hoechs-PIHoechsDNAPI


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3

DNA

DNADNADNADNADNA180-200bpDNADNA


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4(ELISA )

ELISA DNA 5102/m L

:

(1);

(2) ;

(3)


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5DNA

1993WifsmanDNA Fehsel1994TUNEL

:

(1)DNA TUNEL

(2)TUNEL


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Tunel DNA-DNA


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DAB

75 85 95

Tunel


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TunelDAB125


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TunelBCIP/NBT 250


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RNA(RNase)

DNARNA DNARNA mRNA cDNA RNA

32PRNARNA RNA RNA mRNA


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RT-PCR mRNA

RNA RNA(RNA sin) dNTPcDNA cDNAPCR PCR mRNA


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1

2

3

4

5


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40


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1

Switch point gene


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2Killer gene?

3

4

5 survival gene?


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NN


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