家族遗传性胃癌 课件

家族遗传性胃癌:基因诊断、筛查和临床处理贾淑芹北京大学肿瘤医院

分子诊断中心1家族遗传性胃癌:基因诊断、筛查和临床处理贾淑芹1AutosomalDominantInheritedCancer

Syndromes•Breast

and

Ovarian

Cancer

BRCA1&2ColonCancerand

Polyposis2HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile

PolyposisMMRAPCMYHPTENSTK11SMAD4BMPR1AOtherGI

CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16MeninRETVHLp53北京大学肿瘤医院（PKUCH)分子诊断中心(MDC)癌症遗传基因筛查101基因panel为癌症患者及其家系成员进行风险评估、遗传咨询和干预AutosomalDominantInherited2333癌症的遗传易感性4• 谁应该做癌症遗传易感性检测？• 若携带与癌症相关的已知基因突变，患癌的风险有多大？• 对于未发病的携带者，我们能做些什么？癌症的遗传易感性4• 谁应该做癌症遗传易感性检测？• 若携带4•遗传弥漫型胃癌(HDGC)•林奇综合征(Lynch)•遗传性乳腺癌卵巢癌综合征(HBOC)•青少年息肉综合征(JPS

)•黑斑息肉综合征(

PJS)90%•家族性腺瘤息肉病(FAP

)•李-佛美尼综合征（LFS)•……家族遗传性胃癌（-）背景5•遗传弥漫型胃癌(HDGC)家族遗传性胃癌（-）背景55家族性胃癌的遗传风险6Chun&Ford,CancerJ.

2012SyndromeGeneFrequencyGC

riskHDGCCDH11-3%>56-70%Lynch

syndromeMMR1/4406-13%HBOCBRCA1&21/40-1/4002.5-5%Juvenile

PolyposisSMAD4,BMPR1A1/16-100,00021%Peutz-JegherSTK111/25-250,00030%FAPAPC1/10-100,0002-4%Li-FraumeniP531/50003-5%家族性胃癌的遗传风险6Chun&Ford,Cancer6遗传性弥漫型胃癌• 1998年，首次发现于新西兰3个毛利家族中，早发的胃癌显示出常染色体显性遗传模式• 连锁分析将基因定位于16q22.1• 在3个家系中都发现E-cadherin

(CDH-1）种系截短突变• 外显率：60岁前，70%的患者均患癌7遗传性弥漫型胃癌• 1998年，首次发现于新西兰3个毛利家族7遗传性弥漫型胃癌

常染色体显性遗传

早发:

14~69y, 平均:

37yLauren分型:

弥散型

不易早期诊断，预后差• CDH1

种系突变为特征8遗传性弥漫型胃癌 常染色体显性遗传88HDGC

诊断标准（IGCLC)2.3confirmedDGCcasesin1stor2nddegreerelativesindependentofageSingle

DGC<40PersonalorfamilyhistoryofDGCandLBC,1

case<50Single

DGC<40PersonalorfamilyhistoryofDGCandLBC,1case<5020101. 2GCcases

in

family,oneconfirmed

DGC<5020151.2GCcasesinfamily,oneconfirmed

DGC19991.2GCcasesinfamily,oneconfirmed

DGC<502.3confirmedDGCcasesin1stor2nddegreerelativesindependentofage9HDGC诊断标准（IGCLC)2.3confirmed92015

版标准解读10• 2例以上胃癌，至少其中一例是弥漫型~30-40%

种系突变• 年龄<40岁的弥漫型胃癌~10%

种系突变• 任何一个家属同时具有弥漫型胃癌和乳腺小叶癌，其中至少一例<50岁• 个人双侧乳腺小叶癌或者多个家族成员乳腺小叶癌，一例<

50岁• 患者同时具有弥漫型胃癌和唇/腭裂• 印戒细胞癌的癌前病变2015版标准解读10• 2例以上胃癌，至少其中一例是弥漫101111CDH1突变的患者印戒细胞癌中E-cadherin表达降低或缺如12CDH1突变的患者印戒细胞癌中E-cadherin表12CDH1

与患癌风险• 终生患癌风险:男性携带者--70%

GC女性携带者---56%

GC女性携带者---42%

乳腺小叶癌(LBC)• 中位发病年龄

毛利族:32

yrs其他：

43yrs13Hansford&Huntsman,JAMAOnc

2015CDH1与患癌风险• 终生患癌风险:13Hansford• 终生患癌风险:男性携带者--67%

GC女性携带者---83%

GC女性携带者---60%

乳腺小叶癌(LBC)• 18-40ys

携带者建议行预防性全胃切除术14CDH1与患癌风险(2016NCCN,

V1）• 终生患癌风险:男性携带者--67%GC14CDH1HDGC中CDH1种系突变的地域差异15• 低风险区（北美、加拿大、英国）---50%• 中风险区（德国）---25%• 高风险区(葡萄牙，意大利)

---22%• 散发性胃癌高发区(中、日、韩)

~10%HDGC中CDH1种系突变的地域差异15• 低风险区（北美、HDGC中CDH1的突变定位16HDGC中CDH1的突变定位16临床处理遗传学检测和咨询

(E-cadherinorCDH1)• 年龄<25岁或早于先证者10年，每6-12个月做一次胃镜检查，盲取活检30块• 乳腺X光

/乳腺核磁• 预防性全胃切除vanderPost,etal.Journalof

medicalgenetics,

2015.17临床处理遗传学检测和咨询(E-cadherinorCD家族遗传性胃癌（二）我们的研究18家族遗传性胃癌（二）我们的研究18Materialand

Methods19Samples:103

HDGC1500sporadic

GC3097normalcontrolMethods:16exonsPCR+

sequencing~~GC：sitesequencing~~Familiesand

controls：MLPA~~HDGCMaterialandMethods19Samples:VariablesTotalNo.ofpatients(N=1603)Age,

years≦3561>35and

<651051≧65491SexMale1145Female458Lauren

classificationIntestinal517Diffuse864GCfamilial

aggregationPositive182Negative1421HDGC(2010

criteria)82HDGC(2015

criteria)9220Results（1）Table1.Patient

informationVariablesTotalNo.ofpatientsc.1018A>G，p.T340A，7

casesc.865G>A p.A289T，1

casec.1273G>C，p.V425L，1case2.CDH1germline

mutationc.1888C>G，

p.L630V，34casesc.2165-1G>A，

1case21c.1018A>G，p.T340A，7casesc.865c.1298A>Gp.D433G,1

casec.2206G>Ap.V736M，

1casec.1103C>GTp.T368S，1

casec.1174G>Ap.V392I，

1casec.1581A>Cp.R527S，

1case22c.1298A>Gp.D433G,1casec.22061case，exon14

deletion3.CDH1rearrangementin

HDGCNormalE14delE14DELNormal23E14del1case，exon14deletion3.CDH1V736MR527SL630VA289TT340AT368SV392IV425LD433G/N4. SitesofCDH1germline

mutations---------------------------------------------------------------------------------------------Missense

mutation2165-1

G>AE14delRearrangement---------------------------------------------------------------------------------------------Splicesite

mutation24V736MR527SL630VA289T4. Siteso5.CDH1L630Vand

GC

25CDH1L630VmutationinGC

andnormal

controlstotalmutation

no.ratePNormal

control3097571.84%GC1591342.14%0.484HDGC（2010）8244.88%0.071HDGC（2015）9144.40%0.0955.CDH1L630VandGC 25CDH1L256.SummaryExonSitesTypePolymorphismFunctional prediction CasesMutationrateMutation

rateinHDGC in

HDGCMutationReferencep.T340Amissense--benign71/82

=0.0121/92

=0.01106253rsnumber MAF PloyPhen SIFT(2010) (2015) ratein

GC-2affect7 c.865G>A

missense - - possibly proteinp.A289T damaging1---- 1/734=0.0014 0function8 c.1018A>G deleteriou 6/1591=0.0s8 c.1103C>GTp.T368Smissensers367868307NAbenigntolerated1---- 1/734=0.00 0149c.1298A>Gp.D433Gmissensers376097289NApossiblyaffectdamaging

proteinfunction1----1/734=0.00140p.V392Imissensers1418640440.0008benigntolerated1----01401406/92=0.06102119 c.1174G>A 1/734=0.0affect9 c.1273G>C

missense - - possibly proteinp.V425L damaging1---- 1/734=0.00function11 c.1581A>Cp.R527S missense - - benign tolerated1---- 1/734=0.0 0014affect12 c.1888C>G probablyp.L630V missense rs2276331 G=0.0012 damaging protein364/82=0.04334/1568=0.functionaffect14 c.2206G>A probably

protein 1 -- -- 1/734=0.00p.V736M missense - - damaging 14

0function14c.2165-1G>Aspliceprobablydamagingaffectproteinfunction11/82=0.141/92=0.111/734=0.0014014E14delrearrangementaffectproteinfunctionaffectproteinfunction11/82=0.141/92=0.11--0266.SummaryExonSitesTypePolymorp26Results（2）CellfunctionstudyforCDH1

L630V27Results（2）27aminoacid

127154708 731

777882EGFR

activationExtracellular

domainJuxtamemberane

domainSrckinase

activationP38

activationL630V28aminoacid 127154708 73177788BFigure1Confirmationofwild-typeormutantCDH1(L630V)FLAGfusionproteinexpressingingastriccancercelllineandCHOcellline(ANCI-N87celllineBCHOcell

line).AFLAGβ-actin(NCI-N87cell

line)FLAGβ-actin(CHOcell

line)29BFigure1Confirmationofwild1.FunctionofL630VinNCI-N87

cell1.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

NCI-N87cell

line.OD

Value

(490nm)00.60.70.824h48h72h96hMockWTL630V0.50.40.30.20.130Figure2CDH1hadnosignificantdifferenceontheproliferationofNCI-N87gastriccancer

cells,Hours1.FunctionofL630VinNCI-N80%20%40%MockWTL630V24h36h

**

** 31Percentage

of

motile

cells1.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

NCI-N87cell

line.Figure3CDH1mutantL630VpromotedthemigrationofNCI-N87gastriccancercellsthroughwoundhealing

assay.A0%20%40%MockWTL630V24h36h *2.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

CHOcell

line.2.FunctionofL630VinCHOcell

line00.80.60.40.211.21.41.624h48h72h96hMockWTL630V32OD

Value

(490nm)Figure4CDH1hadnosignificantdifferenceontheproliferationofCHOcancer

cells,Hours2.1TheinfluenceofCDH1andMockWT2.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

CHOcell

line.0h 12h 24hFigure5CDH1mutantL630VpromotedthemigrationofCHOcancercells

throughwoundhealing

assay.Percentage

of

motile

cells20%L630V0%40%60%MockWTL630V80%12h24h**33MockWT2.2TheinfluenceofCDHDISCUSSIONCDH1anditsmutantL630Vhadno

significantinfluence

on cancercellproliferation

.CDH1mutantL630Vpromotedthemigration

ofgastriccancer

cells.34DISCUSSION34结 论• 中国人GC中CDH1种系突变以错义突变为主，突变频率为3.14%（50/1591）• HDGC中CDH1突变频率为8.53%（or

9.78%）• CDH1

T340A很可能是HDGC的致病突变• CDH1 L630V在我国正常人群和GC、HDGC人群中突变频率均无显著差异35结 论35结 论36• 90%以上的HDGC

无CDH1种系突变，CDH1种系突变可能不是我国HDGC的主要遗传易感基因。• 本研究为国内家族遗传性胃癌的预防性切除术积累数据，提供理论及实践的基础。结 论36• 90%以上的HDGC无CDH1种系突变，CD家族遗传性胃癌（三）案例分析37家族遗传性胃癌（三）案例分析3735号家系

CDH1T340A:carrier:gastriccancerLungcancerEsophegeal

cancerBreastcancerTestedCancerSomaticmutation

T340A61y3834y40y35号家系CDH1T340A:carrier61y38335号家属，34y，男性，CDH1

T340A3935号家属，34y，男性，CDH1T340A39688号家系

CDH1L630V40GC:gastric

cancerIGC:intestinal

GCEndC:endometrial

cancer688号家系CDH1L630V40GC:gastric688-241688-241688-442688-442688-843688-843688-944688-944688-1445688-1445建议流

程46• 详细地家系咨询，患者知情同意，遗传学检测• 家系随访

“肿瘤易感基因panel”检测及复检• 通知18-40y，无症状致病突变携带者在遗传门诊咨询• 建议进行胃镜检测及取24-30块活检、病检• 根据结果进一步遗传门诊治疗或随访建议建议流程46• 详细地家系咨询，患者知情同意，遗传学检4747家族遗传性胃癌:基因诊断、筛查和临床处理贾淑芹北京大学肿瘤医院

分子诊断中心1家族遗传性胃癌:基因诊断、筛查和临床处理贾淑芹48AutosomalDominantInheritedCancer

Syndromes•Breast

and

Ovarian

Cancer

BRCA1&2ColonCancerand

Polyposis49HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile

PolyposisMMRAPCMYHPTENSTK11SMAD4BMPR1AOtherGI

CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16MeninRETVHLp53北京大学肿瘤医院（PKUCH)分子诊断中心(MDC)癌症遗传基因筛查101基因panel为癌症患者及其家系成员进行风险评估、遗传咨询和干预AutosomalDominantInherited4950350癌症的遗传易感性51• 谁应该做癌症遗传易感性检测？• 若携带与癌症相关的已知基因突变，患癌的风险有多大？• 对于未发病的携带者，我们能做些什么？癌症的遗传易感性4• 谁应该做癌症遗传易感性检测？• 若携带51•遗传弥漫型胃癌(HDGC)•林奇综合征(Lynch)•遗传性乳腺癌卵巢癌综合征(HBOC)•青少年息肉综合征(JPS

)•黑斑息肉综合征(

PJS)90%•家族性腺瘤息肉病(FAP

)•李-佛美尼综合征（LFS)•……家族遗传性胃癌（-）背景52•遗传弥漫型胃癌(HDGC)家族遗传性胃癌（-）背景552家族性胃癌的遗传风险53Chun&Ford,CancerJ.

2012SyndromeGeneFrequencyGC

riskHDGCCDH11-3%>56-70%Lynch

syndromeMMR1/4406-13%HBOCBRCA1&21/40-1/4002.5-5%Juvenile

PolyposisSMAD4,BMPR1A1/16-100,00021%Peutz-JegherSTK111/25-250,00030%FAPAPC1/10-100,0002-4%Li-FraumeniP531/50003-5%家族性胃癌的遗传风险6Chun&Ford,Cancer53遗传性弥漫型胃癌• 1998年，首次发现于新西兰3个毛利家族中，早发的胃癌显示出常染色体显性遗传模式• 连锁分析将基因定位于16q22.1• 在3个家系中都发现E-cadherin

(CDH-1）种系截短突变• 外显率：60岁前，70%的患者均患癌54遗传性弥漫型胃癌• 1998年，首次发现于新西兰3个毛利家族54遗传性弥漫型胃癌

常染色体显性遗传

早发:

14~69y, 平均:

37yLauren分型:

弥散型

不易早期诊断，预后差• CDH1

种系突变为特征55遗传性弥漫型胃癌 常染色体显性遗传855HDGC

诊断标准（IGCLC)2.3confirmedDGCcasesin1stor2nddegreerelativesindependentofageSingle

DGC<40PersonalorfamilyhistoryofDGCandLBC,1

case<50Single

DGC<40PersonalorfamilyhistoryofDGCandLBC,1case<5020101. 2GCcases

in

family,oneconfirmed

DGC<5020151.2GCcasesinfamily,oneconfirmed

DGC19991.2GCcasesinfamily,oneconfirmed

DGC<502.3confirmedDGCcasesin1stor2nddegreerelativesindependentofage56HDGC诊断标准（IGCLC)2.3confirmed562015

版标准解读57• 2例以上胃癌，至少其中一例是弥漫型~30-40%

种系突变• 年龄<40岁的弥漫型胃癌~10%

种系突变• 任何一个家属同时具有弥漫型胃癌和乳腺小叶癌，其中至少一例<50岁• 个人双侧乳腺小叶癌或者多个家族成员乳腺小叶癌，一例<

50岁• 患者同时具有弥漫型胃癌和唇/腭裂• 印戒细胞癌的癌前病变2015版标准解读10• 2例以上胃癌，至少其中一例是弥漫575811CDH1突变的患者印戒细胞癌中E-cadherin表达降低或缺如59CDH1突变的患者印戒细胞癌中E-cadherin表12CDH1

与患癌风险• 终生患癌风险:男性携带者--70%

GC女性携带者---56%

GC女性携带者---42%

乳腺小叶癌(LBC)• 中位发病年龄

毛利族:32

yrs其他：

43yrs60Hansford&Huntsman,JAMAOnc

2015CDH1与患癌风险• 终生患癌风险:13Hansford• 终生患癌风险:男性携带者--67%

GC女性携带者---83%

GC女性携带者---60%

乳腺小叶癌(LBC)• 18-40ys

携带者建议行预防性全胃切除术61CDH1与患癌风险(2016NCCN,

V1）• 终生患癌风险:男性携带者--67%GC14CDH1HDGC中CDH1种系突变的地域差异62• 低风险区（北美、加拿大、英国）---50%• 中风险区（德国）---25%• 高风险区(葡萄牙，意大利)

---22%• 散发性胃癌高发区(中、日、韩)

~10%HDGC中CDH1种系突变的地域差异15• 低风险区（北美、HDGC中CDH1的突变定位63HDGC中CDH1的突变定位16临床处理遗传学检测和咨询

(E-cadherinorCDH1)• 年龄<25岁或早于先证者10年，每6-12个月做一次胃镜检查，盲取活检30块• 乳腺X光

/乳腺核磁• 预防性全胃切除vanderPost,etal.Journalof

medicalgenetics,

2015.64临床处理遗传学检测和咨询(E-cadherinorCD家族遗传性胃癌（二）我们的研究65家族遗传性胃癌（二）我们的研究18Materialand

Methods66Samples:103

HDGC1500sporadic

GC3097normalcontrolMethods:16exonsPCR+

sequencing~~GC：sitesequencing~~Familiesand

controls：MLPA~~HDGCMaterialandMethods19Samples:VariablesTotalNo.ofpatients(N=1603)Age,

years≦3561>35and

<651051≧65491SexMale1145Female458Lauren

classificationIntestinal517Diffuse864GCfamilial

aggregationPositive182Negative1421HDGC(2010

criteria)82HDGC(2015

criteria)9267Results（1）Table1.Patient

informationVariablesTotalNo.ofpatientsc.1018A>G，p.T340A，7

casesc.865G>A p.A289T，1

casec.1273G>C，p.V425L，1case2.CDH1germline

mutationc.1888C>G，

p.L630V，34casesc.2165-1G>A，

1case68c.1018A>G，p.T340A，7casesc.865c.1298A>Gp.D433G,1

casec.2206G>Ap.V736M，

1casec.1103C>GTp.T368S，1

casec.1174G>Ap.V392I，

1casec.1581A>Cp.R527S，

1case69c.1298A>Gp.D433G,1casec.22061case，exon14

deletion3.CDH1rearrangementin

HDGCNormalE14delE14DELNormal70E14del1case，exon14deletion3.CDH1V736MR527SL630VA289TT340AT368SV392IV425LD433G/N4. SitesofCDH1germline

mutations---------------------------------------------------------------------------------------------Missense

mutation2165-1

G>AE14delRearrangement---------------------------------------------------------------------------------------------Splicesite

mutation71V736MR527SL630VA289T4. Siteso5.CDH1L630Vand

GC

72CDH1L630VmutationinGC

andnormal

controlstotalmutation

no.ratePNormal

control3097571.84%GC1591342.14%0.484HDGC（2010）8244.88%0.071HDGC（2015）9144.40%0.0955.CDH1L630VandGC 25CDH1L726.SummaryExonSitesTypePolymorphismFunctional prediction CasesMutationrateMutation

rateinHDGC in

HDGCMutationReferencep.T340Amissense--benign71/82

=0.0121/92

=0.01106253rsnumber MAF PloyPhen SIFT(2010) (2015) ratein

GC-2affect7 c.865G>A

missense - - possibly proteinp.A289T damaging1---- 1/734=0.0014 0function8 c.1018A>G deleteriou 6/1591=0.0s8 c.1103C>GTp.T368Smissensers367868307NAbenigntolerated1---- 1/734=0.00 0149c.1298A>Gp.D433Gmissensers376097289NApossiblyaffectdamaging

proteinfunction1----1/734=0.00140p.V392Imissensers1418640440.0008benigntolerated1----01401406/92=0.06102119 c.1174G>A 1/734=0.0affect9 c.1273G>C

missense - - possibly proteinp.V425L damaging1---- 1/734=0.00function11 c.1581A>Cp.R527S missense - - benign tolerated1---- 1/734=0.0 0014affect12 c.1888C>G probablyp.L630V missense rs2276331 G=0.0012 damaging protein364/82=0.04334/1568=0.functionaffect14 c.2206G>A probably

protein 1 -- -- 1/734=0.00p.V736M missense - - damaging 14

0function14c.2165-1G>Aspliceprobablydamagingaffectproteinfunction11/82=0.141/92=0.111/734=0.0014014E14delrearrangementaffectproteinfunctionaffectproteinfunction11/82=0.141/92=0.11--0266.SummaryExonSitesTypePolymorp73Results（2）CellfunctionstudyforCDH1

L630V74Results（2）27aminoacid

127154708 731

777882EGFR

activationExtracellular

domainJuxtamemberane

domainSrckinase

activationP38

activationL630V75aminoacid 127154708 73177788BFigure1Confirmationofwild-typeormutantCDH1(L630V)FLAGfusionproteinexpressingingastriccancercelllineandCHOcellline(ANCI-N87celllineBCHOcell

line).AFLAGβ-actin(NCI-N87cell

line)FLAGβ-actin(CHOcell

line)76BFigure1Confirmationofwild1.FunctionofL630VinNCI-N87

cell1.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

NCI-N87cell

line.OD

Value

(490nm)00.60.70.824h48h72h96hMockWTL630V0.50.40.30.20.177Figure2CDH1hadnosignificantdifferenceontheproliferationofNCI-N87gastriccancer

cells,Hours1.FunctionofL630VinNCI-N80%20%40%MockWTL630V24h36h

**

** 78Percentage

of

motile

cells1.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

NCI-N87cell

line.Figure3CDH1mutantL630VpromotedthemigrationofNCI-N87gastriccancercellsthroughwoundhealing

assay.A0%20%40%MockWTL630V24h36h *2.1TheinfluenceofCDH1andits