Download - Cell Cycle Checkpoint and Cancer
Cell Cycle Checkpoint and Cell Cycle Checkpoint and CancerCancer
R4 R4 李日翔 李日翔 / VS / VS 林璟宏林璟宏
Introduction and Cell CycleIntroduction and Cell Cycle DNA damage and CheckpointDNA damage and Checkpoint Checkpoint and CarcinogenesisCheckpoint and Carcinogenesis Checkpoint and Drug TherapyCheckpoint and Drug Therapy
Introduction and Cell CycleIntroduction and Cell Cycle
Nature Medicine 2004;10:789-799
Oncogene
Tumor Suppressor
Apoptosis
Cancer predisposing gene
Control of cell cycle by cdk-cyclin complexesControl of cell cycle by cdk-cyclin complexes
Molecular cell biology
S / M phase kinase :
cdk + cyclin
Regulation of Cell CycleRegulation of Cell Cycle
Cell 2004;116:221-34
INK4
(RB)
Cell 2004;116:235-46
DNA damage and DNA damage and checkpointcheckpointTheory of checkpointTheory of checkpointATM proteinATM protein
CheckpointCheckpoint
A control loop that makes the initiation of one A control loop that makes the initiation of one event in the cell cycle dependent on the event in the cell cycle dependent on the successful completion of an earlier eventsuccessful completion of an earlier event A cell must replicate every sequence of DNA A cell must replicate every sequence of DNA
once and only onceonce and only once The mass of the cell must The mass of the cell must doubledouble
Gene VIII p845-6
CheckpointCheckpoint Initiation of late events is dependent on Initiation of late events is dependent on
the completion of early eventsthe completion of early events Substrate-product orderSubstrate-product order
If… then…If… then… Complement cascade, apoptosis cascadeComplement cascade, apoptosis cascade Give enough substrate, and yield productGive enough substrate, and yield product
Control mechanismControl mechanism ……unless…unless… Found some chemical / mutant relieve a Found some chemical / mutant relieve a
dependent relationshipdependent relationship Relief of dependenceRelief of dependence
Hartwell and Weinerg, Science 1989;246:629-633
Checkpoint
Molecular biology of the cell 2002
p53 and checkpointp53 and checkpoint
Molecular cell biology
p53 mutant let DNA damaged cell enter S phase but not impact G2 checkpoint
RAD9 system in yeastRAD9 system in yeast
cdc9 (DNA ligase) mutation cdc9 (DNA ligase) mutation DNA replicat DNA replicated but unligated ed but unligated arrest at S phase arrest at S phase
cdc9-rad9 double mutant:cdc9-rad9 double mutant: Enter into mitosisEnter into mitosis Cell die much more rapidlyCell die much more rapidly Loss chromosome spontaneously 21X than wilLoss chromosome spontaneously 21X than wil
d-typed-type
Science 1989;246:629-633
Molecular cell biology
Damage of checkpoint
Stopped by next
checkpoint
ATM ATM (Ataxia Telangiectasia mutated)(Ataxia Telangiectasia mutated)
Heterozygous for AT mildly manifest cancer Heterozygous for AT mildly manifest cancer predisposition and radiation sensitivitypredisposition and radiation sensitivity
Cell of AT patients have a reduced life-span,Cell of AT patients have a reduced life-span, higher requirements for serum growth facto higher requirements for serum growth factor, defect in cytoskeletonr, defect in cytoskeleton
Inhibition of semiconservative DNA synthesiInhibition of semiconservative DNA synthesis and cell cycle progression by ionizing irrads and cell cycle progression by ionizing irradiation is reduced in AT celliation is reduced in AT cell
Science 1995;26:1749-53
ATMATM
AT cell failed to induced p53 in response AT cell failed to induced p53 in response to irradiationto irradiation ATM act upstream to p53ATM act upstream to p53
Identify ATM gene from patient with ATIdentify ATM gene from patient with AT Similarity between ATM and rad3Similarity between ATM and rad3
rad3 protein is required for the G2-M rad3 protein is required for the G2-M checkpoint, it monitor the completion of DNA checkpoint, it monitor the completion of DNA damage repairdamage repair
Similarity between ATM and PI3-kinaseSimilarity between ATM and PI3-kinaseCell 1992;71:587-95
Science 1995;26:1749-53
TIBS 1995;20:426-30
mTOR, p110…
TIBS 1995;20:426-30
p53p53
Nature 2004;432:316-23
Trends in Cell Biology 2002;12:509-16
Trends in Cell Biology 2002;12:509-16
Cancer cell 2003;3:421-429
ATM
Checkpoint in S. cerevisiaeCheckpoint in S. cerevisiae
Science 2002;297:552-7
DNA damage
Hus1/Rad1/Rad9
Chk1 BRCA1
DNA repairCell cyclearrest
Others
ATM
ATRRad17/RFC
Apoptosis
or
Human DNA damage response pathway
Replication block
p53
Sensors
Effectors
Signals
Transducers
Common intermediate
Nature 2000;408:433-9
Checkpoint and Checkpoint and CarcinogenesisCarcinogenesis
Checkpoint and carcinogenesisCheckpoint and carcinogenesis
Nature 2004;432:316-23
Adenoma-Carcinoma Sequence
Why the patient so unlucky?Why the patient so unlucky? Dose each oncogene mutate randomly anDose each oncogene mutate randomly an
d independently?d independently? Dose tumor progress steadily?Dose tumor progress steadily?
Science 2002;297:565-9
Oncogene induced a ‘oncogenic stress’ (cyclin E), which activate DNA repair response (ATM, pChk2)
Nature 2005;434:864-70
U-2-OS cell
Cyclin E expressed in a Tetracyclin-repressible manner
Mcm7: loading control
Nature 2005;434:864-70
Absent
Low level
Medium level
High level
pT-Chk2 level in bladder cancer
Checkpoint damage in cancerCheckpoint damage in cancer(bladder cancer)(bladder cancer)
Nature 2005;434:864-70
Genome instability during cancer progressionGenome instability during cancer progression
Nature 2005;434:907-13
Nature 2005;434:907-13
TUNEL stain: hallmark of apoptosis
SummarySummary
Oncogene mutationOncogene mutation hyperplasia hyperplasia DNA replication stress DNA replication stress Double strand brea Double strand brea
kk Activate checkpoint Activate checkpoint cell cycle arrest, DNA r cell cycle arrest, DNA r
epair, or apoptosis epair, or apoptosis no tumor progress no tumor progress Defect of checkpoint (ATM-Chk2-p53)Defect of checkpoint (ATM-Chk2-p53) Genomic instability and cancerGenomic instability and cancer
Nature 2005;434:864-70
Nature 2005;434:907-13
A model for carcinogenesisA model for carcinogenesis
Nature 2005;434:907-13
Science 2008;319:1352-5
Science 2008;319:1352-5
P.I.: proliferation index
DDR.I.: DNA damage reponse index
A./S.I.: apoptosis or senescence index
Checkpoint and Drug Checkpoint and Drug TherapyTherapy
Chemotherapy with intact checkpointChemotherapy with intact checkpointChemotherapy with defective checkpointChemotherapy with defective checkpointTargeting checkpointTargeting checkpoint
Mechanism of alkylating agentMechanism of alkylating agent
Cancer Principles and proctice of Oncology, 7ed p339
Therapy with intact checkpointTherapy with intact checkpoint
DNA damage activate DNA repair and DNA damage activate DNA repair and checkpoint pathwaycheckpoint pathway
ATM->Chk1/2->p53->p21ATM->Chk1/2->p53->p21 Cell cycle arrest or death?Cell cycle arrest or death? Therapy shift arrest to deathTherapy shift arrest to death
Arrest after SN-38Arrest after SN-38
(CPT-11)
JCO 2005;23:4499-4508
FlavopiridolFlavopiridol
CDK inhibitor Inhibit bcl-2, XIAP, p21,
mcl-1, cyclin D1,30 and phospho-survivin
Inhibit p21-induced cell cInhibit p21-induced cell cycle arrest at G2 checkpycle arrest at G2 checkpointoint
JCO 2005;23:4499-4508
JCO 2005;23:4499-4508
Therapy with defective checkpointTherapy with defective checkpoint Response to Flavopiridol according to pResponse to Flavopiridol according to p
53 status53 status Resistance to anti-CD33 in AML cell unResistance to anti-CD33 in AML cell un
able to activate Chk1/Chk2 able to activate Chk1/Chk2
JCO 2005;23:4499-4508
Blood 2003;101:4589-97
Targeting checkpointTargeting checkpoint
Loss of ATM sensitized p53-deficient cell to Loss of ATM sensitized p53-deficient cell to topoisomerase poisons and anti-metabolitetopoisomerase poisons and anti-metabolitess
SN-38 followed by UCN-01 (inhibit Chk1 /CSN-38 followed by UCN-01 (inhibit Chk1 /Chk2) enhance apoptosis in cell with mutated hk2) enhance apoptosis in cell with mutated p53 but not intact p53 p53 but not intact p53
Under development: CHIR124, inhibitor of Under development: CHIR124, inhibitor of Chk1, and 17-AAG: suppresses Chk1expreChk1, and 17-AAG: suppresses Chk1expressionssion JCO 2005;23:4499-4508
Annals of Oncology 2003;14:938-45
Flavopiridol inhibit cell cycle arrestFlavopiridol inhibit cell cycle arrestApoptosis
Flavopiridol
Molecular cell biology
Chk1/p21
UCN-01
A model for targeting checkpointA model for targeting checkpoint
JCO 2005;23:4499-4508
So Simple?
Down stream p53 inhibit G1/S checkpoint
Up stream p53 inhibit G2 checkpoint
To be definedTo be defined
What determines DNA repair, What determines DNA repair, cell cycle arrest, or cell cycle arrest, or apoptosis?apoptosis?
Some drug induced cell cycle Some drug induced cell cycle arrest, some apoptosis, and arrest, some apoptosis, and some necrosis?some necrosis?
Different cell line had different Different cell line had different response?response?
Undefined pathway? Undefined pathway? ∆∆p53?p53?
Cancer cell 2003;3:421-429
?Cell 2005;122:21-32
Thanks for your attentionThanks for your attention
Mitotic checkpointMitotic checkpoint
Nature review cancer 2005;5:773-85
Mitotic checkpoint defect in cancerMitotic checkpoint defect in cancer
Nature review cancer 2005;5:773-85
∆∆p53 in checkpointp53 in checkpoint
Cell 2005;122:21-32
∆∆p53 in checkpointp53 in checkpoint
Cell 2005;122:21-32