1、家族遗传性胃家族遗传性胃癌癌:基基因因诊诊断断、筛筛查查和临床处理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&2 Colon Cancer and Polyposis2HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile PolyposisMMR APC MYH PTEN STK11 SMAD4 BMPR1A Other GI CancersGastricPancreasMEN1MEN2/MTCVHLL
2、i-FraumeniCDH1p16 Menin RET VHLp53北京大学肿瘤医院北京大学肿瘤医院（PKUCH)分子诊断中心分子诊断中心(MDC)癌症遗传基因筛查癌症遗传基因筛查101基因基因panel为癌症患者及其家系成员为癌症患者及其家系成员 进行风险评估、进行风险评估、 遗传咨遗传咨询和干预询和干预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？遗传弥漫型胃癌遗传弥漫型胃癌(HDGC)(HDGC)林奇综合征林奇综合征(Lynch)(Lynch)遗传性乳腺癌卵巢癌综合征遗传性乳腺癌卵巢癌综合征(H
3、BOC)(HBOC)青少年息肉综合征青少年息肉综合征( (JPSJPS ) )黑斑息肉综合征黑斑息肉综合征( ( PJS)PJS)90%家族性腺瘤息肉病家族性腺瘤息肉病( ( FAPFAP ) )李李- -佛美尼综合征（佛美尼综合征（LFS)LFS)家族遗传性胃癌（家族遗传性胃癌（- -）背景）背景5家族性胃癌的遗传风险6Chun & Ford, Cancer J. 2012SyndromeGeneFrequencyGC riskHDGCCDH11-3%56-70%Lynch syndromeMMR1/4406-13%HBOCBRCA1&21/40-1/4002.5-5%Juvenile Po
4、lyposisSMAD4,BMPR1A1/16-100,00021%Peutz-JegherSTK111/25-250,00030%FAPAPC1/10-100,0002-4%Li-FraumeniP531/50003-5%遗传性弥漫型胃癌遗传性弥漫型胃癌 19981998年，首次发现于新西兰年，首次发现于新西兰3 3个毛利家族中，早发个毛利家族中，早发 的胃癌显示出常染色体显性遗传模式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连锁分析将基因定位于16q22.116q22.1 在在3 3个家系中都发现个家系中都发现E-cadherinE-cadherin (CDH-1(CDH-1）
5、种系截）种系截 短突变短突变 外显率：外显率：6060岁前，岁前，70%70%的患者均患癌的患者均患癌7遗传性弥漫型胃癌遗传性弥漫型胃癌常染色体显性遗传常染色体显性遗传早发早发: 1469y,平均平均: 37yLauren 分型分型: 弥散型弥散型不易早期诊断，预后差不易早期诊断，预后差 CDH1 种系突变为特征种系突变为特征8HDGCHDGC 诊断标准（诊断标准（IGCLC)IGCLC)2. 3 confirmed DGC cases in 1st or 2nd degree relatives independent of age3.Single DGC404.Personal or fa
6、mily history of DGC and LBC, 1 case502.Single DGC403.Personal or family 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 relative
7、s independent of age92015 版标准解读版标准解读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 与患癌风险与患癌风险 终生患癌风险终生患癌风险:男性携带者男性携带者-70% GC女性携带者女性携带者-56% GC女性携带者女性携带者- 42% 乳腺小叶
8、癌乳腺小叶癌(LBC) 中中位发病位发病年年龄龄 毛利族毛利族: 32 yrs其他：其他： 43 yrs13Hansford & Huntsman, JAMA Onc 2015 终生患癌风险终生患癌风险: 男性携带者男性携带者-67% GC女性携带者女性携带者-83% GC女性携带者女性携带者- 60% 乳腺小叶癌乳腺小叶癌(LBC) 18-40ys 携带者建议行预防性全胃切除术携带者建议行预防性全胃切除术14CDH1 与患癌风险与患癌风险(2016 NCCN, V1）HDGCHDGC中中CDH1CDH1种系突变的地域差异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
9、（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%H HD DG GC C中中C CD DH H1 1的突变定位的突变定位16临床临床处理处理 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.1888CG， p.L630V，34 casesc.2165-1 GA， 1 case21c.1298AGp.D433G, 1 casec.2206GAp.
10、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 of CDH1 germline mutations-Missense mutation2165-1 GAE14delRearrangement-S
11、plice site mutation245. CDH1 L630V and GC 25CDH1 L630V mutation in GC andnormal controlstotalmutation no.raterateP PNormalNormal controlcontrol3097309757571.84%1.84%GCGC1591159134342.14%2.14%0.4840.484HDGCHDGC（20102010）82824 44.88%4.88%0.0710.071HDGCHDGC（20152015）91914 44.40%4.40%0.0950.0956.Summary
12、ExonSitesTypePolymorphismFunctional 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.1018AGdeleter
13、iou6/1591=0.0s8c.1103CGT p.T368S missense rs367868307NAbenigntolerated1-1/734=0.000149c.1298AGp.D433Gmissense rs376097289NApossiblyaffectdamaging proteinfunction1-1/734=0.00140p.V392Imissense rs1418640440.0008benigntolerated1-01401406/92=0.06102119c.1174GA1/734=0.0affect9c.1273GC missense-possiblypr
14、oteinp.V425Ldamaging1-1/734=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 GAspliceproba
15、bly damagingaffect protein 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
16、activationP38 activationL630V28BFigure 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.
17、1 The influence of CDH1 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 o
18、f motile cells1.2 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.A2.1 The influence of CDH1 and its mutant L630V on the proliferation in CHO cell line.2. Fu
19、nction of L630V in CHO 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 CDH1muta
20、nt L630V promoted the 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 on cancer cell proliferation .CDH1mutant L630V promoted the migration ofgastric cancer cells
21、.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 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）案例分析家族遗传性胃癌（三）案例分析3735号家系 CDH1 T340A:carrier:gastric ca
22、ncer Lung cancer Esophegeal cancer Breast cancer TestedCancerSomatic mutation T340A61y3834y40y35号家属，34y，男性，CDH1 T340A39688号家系 CDH1 L630V40GC: gastric cancer IGC: intestinal GCEndC: endometrial cancer688-241688-442688-843688-944688-1445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47
