家族遗传性胃癌课件

1、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1Autosomal Dominant Inherited Cancer SyndromesBreast and Ovarian CancerBRCA1&2Colon Cancer and Polyposis2HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile PolyposisMMR APC MYH PTEN STK11 SMAD4 BMPR1AOther GI CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16 Me

2、nin RET VHLp53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3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？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家族性胃癌的遗传风险6Chun & Ford, Cancer J.

3、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%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

4、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遗传性弥漫型胃癌常染色体显性遗传早发: 1469y,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8HDGC 诊断标准（IGCLC)2. 3 confirmed DGC cases in 1st or 2nd degree relatives independent of ageSingle DGC40Personal or family history of DGC and LBC, 1 case50Single DGC40Personal or family

5、history of DGC and LBC, 1 case5020101.2 GC cases in family, one confirmed DGC5020151. 2 GC cases in family, one confirmed DGC19991. 2 GC cases in family, one confirmed DGC502. 3 confirmed DGC cases in 1st or 2nd degree relatives independent of age92015 版标准解读102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

6、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11CDH1 突变的患者印戒细胞癌中E-cadherin表达降低或缺如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： 43 yrs13Hansford & Huntsman, JAMA Onc 2015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

7、% 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(2016 NCCN, V1）HDGC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的突变定位16临床处理遗传学检测和咨询 (E-cadherin or CDH1)年龄35 and G，p.T340A，7 casesc.865GAp.A289T， 1 casec.1273GC ，p.V425L，1 case2. CDH1 germline mutationc.188

8、8CG， p.L630V，34 casesc.2165-1 GA， 1 case21c.1298AGp.D433G, 1 casec.2206GAp.V736M， 1 casec.1103CGTp.T368S，1 casec.1174GAp.V392I， 1 casec.1581ACp.R527S， 1 case221 case，exon14 deletion3. CDH1 rearrangement in HDGCNormalE14delE14DELNormal23E14delV736MR527SL630VA289TT340A T368S V392I V425LD433G/N4.Sites

9、of CDH1 germline mutations-Missense mutation2165-1 GAE14delRearrangement-Splice site mutation245. CDH1 L630V and GC 25CDH1 L630V mutation in GC andnormal controlstotalmutation no.ratePNormal control3097571.84%GC1591342.14%0.484HDGC（2010）8244.88%0.071HDGC（2015）9144.40%0.0956.SummaryExonSitesTypePolym

10、orphismFunctional predictionCasesMutation rate Mutation ratein HDGCin HDGCMutationReferencep.T340Amissense-benign71/82 =0.0121/92 =0.01106253rs numberMAFPloyPhenSIFT(2010)(2015)rate in GC-2affect7c.865GA missense-possiblyproteinp.A289Tdamaging1-1/734=0.00140function8c.1018AGdeleteriou6/1591=0.0s8c.1

11、103CGT p.T368S missense rs367868307NAbenigntolerated1-1/734=0.000149c.1298AGp.D433Gmissensers376097289NApossiblyaffectdamaging proteinfunction1-1/734=0.00140p.V392Imissensers1418640440.0008benigntolerated1-01401406/92=0.06102119c.1174GA1/734=0.0affect9c.1273GC missense-possiblyproteinp.V425Ldamaging

12、1-1/734=0.00function11c.1581ACp.R527Smissense-benigntolerated1-1/734=0.00014affect12c.1888CGprobablyp.L630Vmissensers2276331G=0.0012damagingprotein364/82=0.04334/1568=0.functionaffect14c.2206GAprobably protein1-1/734=0.00p.V736Mmissense-damaging140function14c.2165-1 GAspliceprobably damagingaffect p

13、rotein function11/82=0.141/92=0.111/734=0.0014014E14delrearrange mentaffect protein functionaffect protein function11/82=0.141/92=0.11-026Results（2）Cell function study for CDH1 L630V27amino acid127154708731 777882EGFR activationExtracellular domainJuxtamemberane domainSrc kinase activationP38 activa

14、tionL630V28BFigure 1 Confirmation of wild-type or mutant CDH1 (L630V)FLAG fusion protein expressing in gastric cancer cell line and CHO cell line (A NCI-N87 cell line B CHO cell line).AFLAG-actin(NCI-N87 cell line)FLAG-actin( CHO cell line)291. Function of L630V in NCI-N87 cell1.1 The influence of C

15、DH1 and its mutant L630V on the proliferation in NCI-N87 cell line.OD Value (490nm)00.60.70.824h48h72h96hMock WTL630V0.50.40.30.20.130Figure 2 CDH1 had no significant difference on the proliferation of NCI-N87 gastric cancer cells,Hours0%20%40%MockWTL630V24h 36h * *31Percentage of motile cells1.2 Th

16、e influence of CDH1 and its mutant L630V on the migration in NCI- N87 cell line.Figure 3 CDH1mutant L630V promoted the migration of NCI-N87 gastric cancer cells through wound healing assay.A2.1 The influence of CDH1 and its mutant L630V on the proliferation in CHO cell line.2. Function of L630V in C

17、HO cell line00.80.60.40.211.21.41.624h48h72h96hMock WT L630V32OD Value (490nm)Figure 4 CDH1 had no significant difference on the proliferation of CHO cancer cells,HoursMockWT2.2 The influence of CDH1 and its mutant L630V on the migration in CHO cell line.0h12h24hFigure 5 CDH1mutant L630V promoted th

18、e migration of CHO cancer cells throughwound healing assay.Percentage of motile cells20%L630V0%40%60%MockWTL630V80%12h24h*33DISCUSSIONCDH1 and its mutant L630V had no significantinfluence oncancer cell proliferation .CDH1mutant L630V promoted the migration ofgastric cancer cells.34结论中国人GC中CDH1种系突变以错义突变为主，突变频 率