家族遺傳性胃癌

1、會(huì)計(jì)學(xué)1家族遺傳性胃癌家族遺傳性胃癌第1頁(yè)/共54頁(yè)Autosomal Dominant Inherited Cancer SyndromesBreast and Ovarian CancerBRCA1&2 Colon Cancer and Polyposis2HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile PolyposisMMR APC MYH PTEN STK11 SMAD4 BMPR1A Other GI CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16 Menin RET

2、 VHLp53第2頁(yè)/共54頁(yè)3第3頁(yè)/共54頁(yè)4 誰(shuí)應(yīng)該做癌癥遺傳易感性檢測(cè)？ 若攜帶與癌癥相關(guān)的已知基因突變，患癌的風(fēng)險(xiǎn)有多大？ 對(duì)于未發(fā)病的攜帶者，我們能做些什么？第4頁(yè)/共54頁(yè)遺傳彌漫型胃癌遺傳彌漫型胃癌(HDGC)(HDGC)林奇綜合征林奇綜合征(Lynch)(Lynch)遺傳性乳腺癌卵巢癌綜合征遺傳性乳腺癌卵巢癌綜合征(HBOC)(HBOC)青少年息肉綜合征青少年息肉綜合征( (JPSJPS ) )黑斑息肉綜合征黑斑息肉綜合征( ( PJS)PJS)90%家族性腺瘤息肉病家族性腺瘤息肉病( ( FAPFAP ) )李李- -佛美尼綜合征（佛美尼綜合征（LFS)LFS)5第5頁(yè)/共

3、54頁(yè)6Chun & Ford, Cancer J. 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%第6頁(yè)/共54頁(yè) 19981998年，首次發(fā)現(xiàn)于新西蘭年，首次發(fā)現(xiàn)于新西蘭3

4、 3個(gè)毛利家族中，早發(fā)個(gè)毛利家族中，早發(fā) 的胃癌顯示出常染色體顯性遺傳模式的胃癌顯示出常染色體顯性遺傳模式 連鎖分析將基因定位于連鎖分析將基因定位于16q22.116q22.1 在在3 3個(gè)家系中都發(fā)現(xiàn)個(gè)家系中都發(fā)現(xiàn)E-cadherinE-cadherin (CDH-1(CDH-1）種系截）種系截 短突變短突變 外顯率：外顯率：6060歲前，歲前，70%70%的患者均患癌的患者均患癌7第7頁(yè)/共54頁(yè)8第8頁(yè)/共54頁(yè)2. 3 confirmed DGC cases in 1st or 2nd degree relatives independent of age3.Single DGC404

5、.Personal or family 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

6、degree relatives independent of age9第9頁(yè)/共54頁(yè)10 2例以上胃癌，至少其中一例是彌漫型30 - 40% 種系突變 年齡40歲的彌漫型胃癌10% 種系突變 任何一個(gè)家屬同時(shí)具有彌漫型胃癌和乳腺小葉癌，其中至 少一例50歲 個(gè)人雙側(cè)乳腺小葉癌或者多個(gè)家族成員乳腺小葉癌，一例 50歲 患者同時(shí)具有彌漫型胃癌和唇/腭裂 印戒細(xì)胞癌的癌前病變第10頁(yè)/共54頁(yè)11第11頁(yè)/共54頁(yè)CDH1 突變的患者印戒細(xì)胞癌中E-cadherin表達(dá)降低或缺如12第12頁(yè)/共54頁(yè) 終生患癌風(fēng)險(xiǎn)終生患癌風(fēng)險(xiǎn):男性攜帶者男性攜帶者-70% GC女性攜帶者女性攜帶者-56% GC

7、女性攜帶者女性攜帶者- 42% 乳腺小葉癌乳腺小葉癌(LBC) 中中位發(fā)病位發(fā)病年年齡齡 毛利族毛利族: 32 yrs其他：其他： 43 yrs13Hansford & Huntsman, JAMA Onc 2015第13頁(yè)/共54頁(yè) 終生患癌風(fēng)險(xiǎn)終生患癌風(fēng)險(xiǎn): 男性攜帶者男性攜帶者-67% GC女性攜帶者女性攜帶者-83% GC女性攜帶者女性攜帶者- 60% 乳腺小葉癌乳腺小葉癌(LBC) 18-40ys 攜帶者建議行預(yù)防性全胃切除術(shù)攜帶者建議行預(yù)防性全胃切除術(shù)14第14頁(yè)/共54頁(yè)15 低風(fēng)險(xiǎn)區(qū)（北美、加拿大、英國(guó)）-50% 中風(fēng)險(xiǎn)區(qū)（德國(guó)）-25% 高風(fēng)險(xiǎn)區(qū)(葡萄牙，意大利) -

8、22% 散發(fā)性胃癌高發(fā)區(qū)(中、日、韓) 10%第15頁(yè)/共54頁(yè)16第16頁(yè)/共54頁(yè) 遺傳學(xué)檢測(cè)和咨詢 (E-cadherin or CDH1)年齡35 and G，p.T340A，7 casesc.865GAp.A289T， 1 casec.1273GC ，p.V425L，1 casec.1888CG， p.L630V，34 casesc.2165-1 GA， 1 case21第21頁(yè)/共54頁(yè)c.1298AGc.2206GAp.V736M， 1 casec.1103CGTp.T368S，1 casec.1174GAp.V392I， 1 casec.1581ACp.R527S， 1 cas

9、e22第22頁(yè)/共54頁(yè)1 case，exon14 deletionNormalE14delE14DELNormal23E14del第23頁(yè)/共54頁(yè)V736MR527SL630VA289TT340A T368S V392I V425LD433G/N-Missense mutation2165-1 GAE14delRearrangement-Splice site mutation24第24頁(yè)/共54頁(yè) 25CDH1 L630V mutation in GC andnormal controlstotalmutation no.raterateP PNormalNormal controlco

10、ntrol3097309757571.84%1.84%GCGC1591159134342.14%2.14%0.4840.484HDGCHDGC（20102010）82824 44.88%4.88%0.0710.071HDGCHDGC（20152015）91914 44.40%4.40%0.0950.095第25頁(yè)/共54頁(yè)ExonSitesTypePolymorphismFunctional predictionCasesMutation rate Mutation ratein HDGCin HDGCMutationReferencep.T340Amissense-benign71/82 =

11、0.0121/92 =0.01106253rs numberMAFPloyPhenSIFT(2010)(2015)rate in GC-2affect7c.865GA missense-possiblyproteinp.A289Tdamaging1-1/734=0.00140function8c.1018AGdeleteriou6/1591=0.0s8c.1103CGT p.T368S missense rs367868307NAbenigntolerated1-1/734=0.000149c.1298AGp.D433Gmissense rs376097289NApossiblyaffectd

12、amaging proteinfunction1-1/734=0.00140p.V392Imissense rs1418640440.0008benigntolerated1-01401406/92=0.06102119c.1174GA1/734=0.0affect9c.1273GC missense-possiblyproteinp.V425Ldamaging1-1/734=0.00function11c.1581ACp.R527Smissense-benigntolerated1-1/734=0.00014affect12c.1888CGprobablyp.L630Vmissensers2

13、276331G=0.0012damagingprotein364/82=0.04334/1568=0.functionaffect14c.2206GAprobably protein1-1/734=0.00p.V736Mmissense-damaging140function14c.2165-1 GAspliceprobably damagingaffect protein function11/82=0.141/92=0.111/734=0.0014014E14delrearrange mentaffect protein functionaffect protein function11/

14、82=0.141/92=0.11-026第26頁(yè)/共54頁(yè)Results（2）Cell function study for CDH1 L630V27第27頁(yè)/共54頁(yè)amino acid127154708731 777882EGFR activationExtracellular domainJuxtamemberane domainSrc kinase activationP38 activationL630V28第28頁(yè)/共54頁(yè)BFigure 1 Confirmation of wild-type or mutant CDH1 (L630V)FLAG fusion protein ex

15、pressing 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)29第29頁(yè)/共54頁(yè)1.1 The influence of CDH1 and its mutant L630V on the proliferation in NCI-N87 cell line.OD Value (490nm)00.60.70.824h48h72h96hMock WTL630V0

16、.50.40.30.20.130Figure 2 CDH1 had no significant difference on the proliferation of NCI-N87 gastric cancer cells,Hours第30頁(yè)/共54頁(yè)0%20%40%MockWTL630V24h 36h * *31Percentage of motile cellsFigure 3 CDH1mutant L630V promoted the migration of NCI-N87 gastric cancer cells through wound healing assay.A第31頁(yè)/

17、共54頁(yè)2.1 The influence of CDH1 and its mutant L630V on the proliferation in CHO cell line.00.80.60.40.211.21.41.624h48h72h96hMock WT L630V32OD Value (490nm)Figure 4 CDH1 had no significant difference on the proliferation of CHO cancer cells,Hours第32頁(yè)/共54頁(yè)MockWTFigure 5 CDH1mutant L630V promoted the m

18、igration of CHO cancer cells throughwound healing assay.Percentage of motile cells20%L630V0%40%60%MockWTL630V80%12h24h*33第33頁(yè)/共54頁(yè)DISCUSSIONCDH1 and its mutant L630V had no significantinfluence on cancer cell proliferation .CDH1mutant L630V promoted the migration ofgastric cancer cells.34第34頁(yè)/共54頁(yè)結(jié)論

19、 中國(guó)人GC中CDH1種系突變以錯(cuò)義突變?yōu)橹?，突變頻 率為3.14%（50/1591） HDGC中CDH1突變頻率為8.53%（or 9.78%） CDH1 T340A很可能是HDGC的致病突變 CDH1 L630V在我國(guó)正常人群和GC、HDGC人群中突 變頻率均無(wú)顯著差異35第35頁(yè)/共54頁(yè)結(jié)論36系突變可能不是我國(guó)HDGC的主要遺傳易感基因。 本研究為國(guó)內(nèi)家族遺傳性胃癌的預(yù)防性切除術(shù)積累數(shù)據(jù)，提供理論及實(shí)踐的基礎(chǔ)。第36頁(yè)/共54頁(yè)37第37頁(yè)/共54頁(yè)35號(hào)家系 CDH1 T340A:carrier:gastric cancer Lung cancer Esophegeal cance

20、r Breast cancer TestedCancerSomatic mutation T340A61y3834y40y第38頁(yè)/共54頁(yè)39第39頁(yè)/共54頁(yè)688號(hào)家系 CDH1 L630V40GC: gastric cancer IGC: intestinal GCEndC: endometrial cancer第40頁(yè)/共54頁(yè)41第41頁(yè)/共54頁(yè)42第42頁(yè)/共54頁(yè)43第43頁(yè)/共54頁(yè)44第44頁(yè)/共54頁(yè)45第45頁(yè)/共54頁(yè)46第46頁(yè)/共54頁(yè)47第47頁(yè)/共54頁(yè)Autosomal Dominant Inherited Cancer SyndromesBreast a

21、nd Ovarian CancerBRCA1&2 Colon Cancer and Polyposis48HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile PolyposisMMR APC MYH PTEN STK11 SMAD4 BMPR1A Other GI CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16 Menin RET VHLp53第48頁(yè)/共54頁(yè)49Samples: 103 HDGC1500 sporadic GC3097 normal controlMethods: 16 exons PCR+ sequencingGC：site sequencingFamilies and control