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1,Autosomal Dominant Inherited Cancer SyndromesBreast and Ovarian CancerBRCA1&2Colon Cancer and Polyposis,2,HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile Polyposis,MMR APC MYH PTEN STK11 SMAD4 BMPR1A,Other GI Cancers,GastricPancreasMEN1MEN2/MTCVHLLi-Fraumeni,
2、CDH1p16 Menin 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,6
3、,Chun & Ford, Cancer J. 2012,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,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,8,HDGC 诊断标准（IGCLC),2. 3 confirmed DGC cases in 1st or 2nd degree relatives ind
4、ependent of age,Single DGC40Personal or family history of DGC and LBC, 1 case50,Single DGC40Personal or family history of DGC and LBC, 1 case50,20101.2 GC cases in family, one confirmed DGC50,20151. 2 GC cases in family, one confirmed DGC,19991. 2 GC cases in family, one confirmed DGC50,2. 3 confirm
5、ed DGC cases in 1st or 2nd degree relatives independent of age,9,2015 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,11,CDH1 突变的患者印戒细胞癌中E-cadherin表达降低或缺如,12,CDH1 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
6、携带者- 42% 乳腺小叶癌(LBC)中位发病年龄 毛利族: 32 yrs其他： 43 yrs,13,Hansford & 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% 乳腺小叶癌(LBC)18-40ys 携带者建议行预防性全胃切除术,14,CDH1 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,
7、临床处理,遗传学检测和咨询 (E-cadherin or CDH1)年龄G，p.T340A，7 cases,c.865GAp.A289T， 1 case,c.1273GC ，p.V425L，1 case,2. CDH1 germline mutation,c.1888CG， p.L630V，34 cases,c.2165-1 GA， 1 case,21,c.1298AG,p.D433G, 1 case,c.2206GA,p.V736M， 1 case,c.1103CGT,p.T368S，1 case,c.1174GA,p.V392I， 1 case,c.1581AC,p.R527S， 1 ca
8、se,22,1 case，exon14 deletion,3. CDH1 rearrangement in HDGC,Normal,E14del,E14DEL,Normal,23,E14del,V736M,R527S,L630V,A289TT340A T368S V392I V425LD433G/N,4.Sites of CDH1 germline mutations,-Missense mutation,2165-1 GA,E14del,Rearrangement-Splice site mutation,24,5. CDH1 L630V and GC,25,CDH1 L630V mutat
9、ion in GC andnormal controls,6.Summary,Exon,Sites,Type,Polymorphism,Functional,prediction,Cases,Mutation rate Mutation ratein HDGCin HDGC,Mutation,Reference,p.T340A,missense,-,-,benign,7,1/82 =0.012,1/92 =0.011,0625,3,9,c.1298AGp.D433G,missense,rs376097289,NA,possibly,affect,damaging protein,functio
10、n,1,-,-,1/734=0.0014,0,p.V392I,missense,rs141864044,0.0008,benign,tolerated,1,-,-,014,0,14,0,6/92=0.061,021,1,14,c.2165-1 GA,splice,probably damaging,affect protein function,1,1/82=0.14,1/92=0.11,1/734=0.0014,0,14,E14del,rearrange ment,affect protein function,affect protein function,1,1/82=0.14,1/92
11、=0.11,-,0,26,Results（2）Cell function study for CDH1 L630V,27,amino acid1,27,154,708731 777,882,EGFR activation,Extracellular domain,Juxtamemberane domain,Src kinase activation,P38 activation,L630V,28,B,Figure 1 Confirmation of wild-type or mutant CDH1 (L630V)FLAG fusion protein expressing in gastric
12、 cancer cell line and CHO cell line (A NCI-N87 cell line B CHO cell line).,A,FLAG,-actin,(NCI-N87 cell line),FLAG,-actin,( CHO cell line),29,1. Function of L630V in NCI-N87 cell,1.1 The influence of CDH1 and its mutant L630V on the proliferation in NCI-N87 cell line.,OD Value (490nm),0,0.6,0.7,0.8,2
13、4h,48h,72h,96h,Mock WT,L630V0.50.40.30.20.1,30,Figure 2 CDH1 had no significant difference on the proliferation of NCI-N87 gastric cancer cells,Hours,0%,20%,40%,Mock,WT,L630V,24h 36h,*,*,31,Percentage of motile cells,1.2 The influence of CDH1 and its mutant L630V on the migration in NCI- N87 cell li
14、ne.,Figure 3 CDH1mutant L630V promoted the migration of NCI-N87 gastric cancer cells through wound healing assay.,A,2.1 The influence of CDH1 and its mutant L630V on the proliferation in CHO cell line.,2. Function of L630V in CHO cell line,0,0.80.60.40.2,1,1.2,1.4,1.6,24h,48h,72h,96h,Mock WT L630V,3
15、2,OD Value (490nm),Figure 4 CDH1 had no significant difference on the proliferation of CHO cancer cells,Hours,Mock,WT,2.2 The influence of CDH1 and its mutant L630V on the migration in CHO cell line.0h12h24h,Figure 5 CDH1mutant L630V promoted the migration of CHO cancer cells throughwound healing as
16、say.,Percentage of motile cells,20%L630V0%,40%,60%,Mock,WT,L630V,80%12h,24h,*,33,DISCUSSIONCDH1 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 率为3.14%（50/1591）HDGC中CDH1突变频率为8
17、.53%（or 9.78%）CDH1 T340A很可能是HDGC的致病突变CDH1L630V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,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。,家族遗传性胃癌（三）案例分析,37,35号家系 CDH1 T340A,:carrier:gastric cancer Lung cancer Esophegeal cancer Breast cancer TestedCancerSomatic mutat
18、ion T340A,61y,38,34y,40y,35号家属，34y，男性，CDH1 T340A,39,688号家系 CDH1 L630V,40,GC: gastric cancer IGC: intestinal GCEndC: endometrial cancer,688-2,41,688-4,42,688-8,43,688-9,44,688-14,45,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,47,
