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Cell Cycle Checkpoint and Cancer

R4 李日翔 / VS 林璟宏

 Introduction and Cell Cycle  DNA damage and Checkpoint  Checkpoint and Carcinogenesis  Checkpoint and Drug Therapy

Introduction and Cell Cycle

Cancer predisposing gene

Tumor Suppressor Apoptosis

Nature Medicine 2004;10:789-799

Control of cell cycle by cdk-cyclin complexes

S / M phase kinase : cdk + cyclin
Molecular cell biology

Regulation of Cell Cycle INK4 (RB) Cell 2004.116:221-34 .

116:235-46 .Cell 2004.

DNA damage and checkpoint Theory of checkpoint ATM protein .

Checkpoint  A control loop that makes the initiation of one event in the cell cycle dependent on the successful completion of an earlier event  A cell must replicate every sequence of DNA once and only once  The mass of the cell must double Gene VIII p845-6 .

and yield product  Control mechanism  Checkpoint  …unless…  Found some chemical / mutant relieve a dependent relationship  Relief of dependence Hartwell and Weinerg.246:629-633 .Checkpoint  Initiation of late events is dependent on the completion of early events  Substrate-product order  If… then…  Complement cascade. Science 1989. apoptosis cascade  Give enough substrate.

Molecular biology of the cell 2002 .

p53 and checkpoint p53 mutant let DNA damaged cell enter S phase but not impact G2 checkpoint Molecular cell biology .

RAD9 system in yeast  cdc9 (DNA ligase) mutation  DNA replicated but unligated  arrest at S phase  cdc9-rad9 double mutant:  Enter into mitosis  Cell die much more rapidly  Loss chromosome spontaneously 21X than wild-type Science 1989.246:629-633 .

Stopped by next checkpoint Damage of checkpoint Molecular cell biology .

higher requirements for serum growth factor. defect in cytoskeleton  Inhibition of semiconservative DNA synthesis and cell cycle progression by ionizing irradiation is reduced in AT cell Science 1995.26:1749-53 .ATM (Ataxia Telangiectasia mutated)  Heterozygous for AT mildly manifest cancer predisposition and radiation sensitivity  Cell of AT patients have a reduced life-span.

it monitor the completion of DNA damage repair  Similarity between ATM and PI3-kinase Cell 1992.ATM  AT cell failed to induced p53 in response to irradiation  ATM act upstream to p53  Identify ATM gene from patient with AT  Similarity between ATM and rad3  rad3 protein is required for the G2-M checkpoint.26:1749-53 .71:587-95 Science 1995.

mTOR. p110… TIBS 1995.20:426-30 .

p53 p53 TIBS 1995.20:426-30 .

432:316-23 .Nature 2004.

Trends in Cell Biology 2002.12:509-16 .

Trends in Cell Biology 2002.12:509-16 .

ATM Cancer cell 2003.3:421-429 .

Checkpoint in S. cerevisiae Science 2002.297:552-7 .

Human DNA damage response pathway DNA damage Replication block Signals Common intermediate ATM or ATR Rad17/RFC Sensors Hus1/Rad1/Rad9 Transducers Chk1 BRCA1 Others p53 Effectors Cell cycle Apoptosis arrest DNA repair .

Nature 2000.408:433-9 .

Checkpoint and Carcinogenesis .

432:316-23 .Checkpoint and carcinogenesis Nature 2004.

Adenoma-Carcinoma Sequence .

 Why the patient so unlucky?  Dose each oncogene mutate randomly and independently?  Dose tumor progress steadily? .

297:565-9 .Science 2002.

pChk2) U-2-OS cell  Cyclin E expressed in a Tetracyclin-repressible manner Mcm7: loading control Nature 2005.Oncogene induced a ‘oncogenic stress’ (cyclin E).434:864-70 . which activate DNA repair response (ATM.

pT-Chk2 level in bladder cancer Absent Low level Medium level High level Nature 2005.434:864-70 .

434:864-70 .Checkpoint damage in cancer (bladder cancer) Nature 2005.

Genome instability during cancer progression Nature 2005.434:907-13 .

TUNEL stain: hallmark of apoptosis Nature 2005.434:907-13 .

or apoptosis  no tumor progress  Defect of checkpoint (ATM-Chk2-p53)  Genomic instability and cancer Nature 2005.434:907-13 .Summary  Oncogene mutation hyperplasia  DNA replication stress  Double strand break  Activate checkpoint  cell cycle arrest. DNA repair.434:864-70 Nature 2005.

434:907-13 .A model for carcinogenesis Nature 2005.

319:1352-5 .Science 2008.

I.: proliferation index DDR.: DNA damage reponse index A.I.319:1352-5 .P.I./S.: apoptosis or senescence index Science 2008.

Checkpoint and Drug Therapy Chemotherapy with intact checkpoint Chemotherapy with defective checkpoint Targeting checkpoint .

Mechanism of alkylating agent Cancer Principles and proctice of Oncology. 7ed p339 .

Therapy with intact checkpoint  DNA damage activate DNA repair and checkpoint pathway  ATM->Chk1/2->p53->p21  Cell cycle arrest or death?  Therapy shift arrest to death .

23:4499-4508 .Arrest after SN-38 (CPT-11) JCO 2005.

Flavopiridol  CDK inhibitor  Inhibit bcl-2. mcl-1.30 and phospho-survivin  Inhibit p21-induced cell cycle arrest at G2 checkpoint JCO 2005. p21.23:4499-4508 . cyclin D1. XIAP.

JCO 2005.23:4499-4508 .

101:4589-97 .23:4499-4508 Blood 2003.Therapy with defective checkpoint  Response to Flavopiridol according to p53 status  Resistance to anti-CD33 in AML cell unable to activate Chk1/Chk2 JCO 2005.

Targeting checkpoint  Loss of ATM sensitized p53-deficient cell to topoisomerase poisons and antimetabolites Annals of Oncology 2003. inhibitor of Chk1.23:4499-4508 .14:938-45  SN-38 followed by UCN-01 (inhibit Chk1 /Chk2) enhance apoptosis in cell with mutated p53 but not intact p53  Under development: CHIR124. and 17-AAG: suppresses Chk1expression JCO 2005.

Flavopiridol inhibit cell cycle arrest Apoptosis Chk1/p21 UCN-01 Flavopiridol Molecular cell biology .

23:4499-4508 .A model for targeting checkpoint Up stream p53 inhibit G2 checkpoint Down stream p53 inhibit G1/S checkpoint So Simple? JCO 2005.

122:21-32 ? Cancer cell 2003.3:421-429 . cell cycle arrest.To be defined  What determines DNA repair. or apoptosis?  Some drug induced cell cycle arrest. some apoptosis. and some necrosis?  Different cell line had different response?  Undefined pathway? ∆p53? Cell 2005.

Thanks for your attention .

Mitotic checkpoint Nature review cancer 2005.5:773-85 .

5:773-85 .Mitotic checkpoint defect in cancer Nature review cancer 2005.

∆p53 in checkpoint Cell 2005.122:21-32 .

122:21-32 .∆p53 in checkpoint Cell 2005.