家族遗传性胃癌ppt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 1,1,Autosomal Dominant Inherited Cancer Syndromes Breast and Ovarian CancerBRCA1&2 Colon Cancer and Polyposis,2,HNPCC FAP Polyposis Cowdens Peutz-Jehgers Juvenile Polyposis,MMR APC MYH PTEN STK11 SMAD4 BMPR1A,Other GI Cancers,Gastric Pancreas MEN1 MEN2/MTC

2、VHL Li-Fraumeni,CDH1 p16 Menin RET VHL p53,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,2,3,3,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？,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

3、尼综合征（LFS) ,家族遗传性胃癌（-）背景,5,5,家族性胃癌的遗传风险,6,Chun & Ford, Cancer J. 2012,6,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,7,7,遗传性弥漫型胃癌,常染色体显性遗传 早发: 1469y,平均: 37y Lauren 分型: 弥散型 不易早期诊断，预后差 CDH1 种系突变为特征,8,8,HDGC 诊断标准（IGCLC),2. 3 c

4、onfirmed DGC cases in 1st or 2nd degree relatives independent of age,Single DGC40 Personal or family history of DGC and LBC, 1 case50,Single DGC40 Personal or family history of DGC and LBC, 1 case50,2010 1.2 GC cases in family, one confirmed DGC50,2015 1. 2 GC cases in family, one confirmed DGC,1999

5、 1. 2 GC cases in family, one confirmed DGC50,2. 3 confirmed DGC cases in 1st or 2nd degree relatives independent of age,9,9,2015 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,10,11,CDH1 突变的患者印

6、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、

7、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,van der Post, et al. Journal of medical genetics, 2015.,17,家族遗传性胃癌（二）我们的研究,18,Material and Methods,19,Samples: 103 HDGC 1500

8、 sporadic GC 3097 normal control Methods: 16 exons PCR+ sequencingGC： site sequencingFamilies and controls： MLPAHDGC,20,Results（1） Table 1. Patient information,c.1018AG，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

9、 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 case,22,1 case，exon14 deletion,3. CDH1 rearrangement in HDGC,Normal,E14del,E14DEL,Normal,23,E14del,V736M,R527S,L630V,A289T T340A T368S V392I V425L D433G/N,4.Sites of CDH1

10、germline mutations,- Missense mutation,2165-1 GA,E14del,Rearrangement - Splice site mutation,24,5. CDH1 L630V and GC,25,CDH1 L630V mutation in GC and normal controls,25,6.Summary,Exon,Sites,Type,Polymorphism,Functional,prediction,Cases,Mutation rate Mutation rate in HDGCin HDGC,Mutation,Reference,p.

11、T340A,missense,-,-,benign,7,1/82 =0.012,1/92 =0.011,0625,3,9,c.1298AG p.D433G,missense,rs376097289,NA,possibly,affect,damaging protein,function,1,-,-,1/734=0.00 14,0,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

12、protein function,1,1/82=0.14,1/92=0.11,1/734=0.0 014,0,14,E14del,rearrange ment,affect protein function,affect protein function,1,1/82=0.14,1/92=0.11,-,0,26,26,Results（2） Cell function study for CDH1 L630V,27,amino acid1,27,154,708731 777,882,EGFR activation,Extracellular domain,Juxtamemberane domai

13、n,Src kinase activation,P38 activation,L630V,28,B,Figure 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).,A,FLAG,-actin,(NCI-N87 cell line),FLAG,-actin,( CHO cell line),29,1. Function

14、 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),0,0.6,0.7,0.8,24h,48h,72h,96h,Mock WT,L630V 0.5 0.4 0.3 0.2 0.1,30,Figure 2 CDH1 had no significant difference on the proliferation of NCI-N87 gastric cancer cells,Hou

15、rs,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 line.,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 i

16、ts mutant L630V on the proliferation in CHO cell line.,2. Function of L630V in CHO cell line,0,0.8 0.6 0.4 0.2,1,1.2,1.4,1.6,24h,48h,72h,96h,Mock WT L630V,32,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

17、and its mutant L630V on the migration in CHO cell line. 0h12h24h,Figure 5 CDH1mutant L630V promoted the migration of CHO cancer cells through wound healing assay.,Percentage of motile cells,20% L630V 0%,40%,60%,Mock,WT,L630V,80% 12h,24h,*,33,DISCUSSION CDH1 and its mutant L630V had no significant in

18、fluence oncancer cell proliferation . CDH1mutant L630V promoted the migration of gastric 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8.53%（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 Tested Cancer Somatic mutation T340A,61y,38