家族遺傳性胃癌 課件

家族遺傳性胃癌:基因診斷、篩查和臨床處理賈淑芹北京大學(xué)腫瘤醫(yī)院

分子診斷中心1家族遺傳性胃癌:基因診斷、篩查和臨床處理賈淑芹1AutosomalDominantInheritedCancer

Syndromes?Breast

and

Ovarian

Cancer

BRCA1&2ColonCancerand

Polyposis2HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile

PolyposisMMRAPCMYHPTENSTK11SMAD4BMPR1AOtherGI

CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16MeninRETVHLp53北京大學(xué)腫瘤醫(yī)院（PKUCH)分子診斷中心(MDC)癌癥遺傳基因篩查101基因panel為癌癥患者及其家系成員進(jìn)行風(fēng)險(xiǎn)評估、遺傳咨詢和干預(yù)AutosomalDominantInherited2333癌癥的遺傳易感性4? 誰應(yīng)該做癌癥遺傳易感性檢測？? 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？? 對于未發(fā)病的攜帶者，我們能做些什么？癌癥的遺傳易感性4? 誰應(yīng)該做癌癥遺傳易感性檢測？? 若攜帶4?遺傳彌漫型胃癌(HDGC)?林奇綜合征(Lynch)?遺傳性乳腺癌卵巢癌綜合征(HBOC)?青少年息肉綜合征(JPS

)?黑斑息肉綜合征(

PJS)90%?家族性腺瘤息肉病(FAP

)?李-佛美尼綜合征（LFS)?……家族遺傳性胃癌（-）背景5?遺傳彌漫型胃癌(HDGC)家族遺傳性胃癌（-）背景55家族性胃癌的遺傳風(fēng)險(xiǎn)6Chun&Ford,CancerJ.

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%家族性胃癌的遺傳風(fēng)險(xiǎn)6Chun&Ford,Cancer6遺傳性彌漫型胃癌? 1998年，首次發(fā)現(xiàn)于新西蘭3個毛利家族中，早發(fā)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16q22.1? 在3個家系中都發(fā)現(xiàn)E-cadherin

(CDH-1）種系截短突變? 外顯率：60歲前，70%的患者均患癌7遺傳性彌漫型胃癌? 1998年，首次發(fā)現(xiàn)于新西蘭3個毛利家族7遺傳性彌漫型胃癌

常染色體顯性遺傳

早發(fā):

14~69y, 平均:

37yLauren分型:

彌散型

不易早期診斷，預(yù)后差? CDH1

種系突變?yōu)樘卣?遺傳性彌漫型胃癌 常染色體顯性遺傳88HDGC

診斷標(biāo)準(zhǔn)（IGCLC)2.3confirmedDGCcasesin1stor2nddegreerelativesindependentofageSingle

DGC<40PersonalorfamilyhistoryofDGCandLBC,1

case<50Single

DGC<40PersonalorfamilyhistoryofDGCandLBC,1case<5020101. 2GCcases

in

family,oneconfirmed

DGC<5020151.2GCcasesinfamily,oneconfirmed

DGC19991.2GCcasesinfamily,oneconfirmed

DGC<502.3confirmedDGCcasesin1stor2nddegreerelativesindependentofage9HDGC診斷標(biāo)準(zhǔn)（IGCLC)2.3confirmed92015

版標(biāo)準(zhǔn)解讀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? 任何一個家屬同時具有彌漫型胃癌和乳腺小葉癌，其中至少一例<50歲? 個人雙側(cè)乳腺小葉癌或者多個家族成員乳腺小葉癌，一例<

50歲? 患者同時具有彌漫型胃癌和唇/腭裂? 印戒細(xì)胞癌的癌前病變2015版標(biāo)準(zhǔn)解讀10? 2例以上胃癌，至少其中一例是彌漫101111CDH1突變的患者印戒細(xì)胞癌中E-cadherin表達(dá)降低或缺如12CDH1突變的患者印戒細(xì)胞癌中E-cadherin表12CDH1

與患癌風(fēng)險(xiǎn)? 終生患癌風(fēng)險(xiǎn):男性攜帶者--70%

GC女性攜帶者---56%

GC女性攜帶者---42%

乳腺小葉癌(LBC)? 中位發(fā)病年齡

毛利族:32

yrs其他：

43yrs13Hansford&Huntsman,JAMAOnc

2015CDH1與患癌風(fēng)險(xiǎn)? 終生患癌風(fēng)險(xiǎn):13Hansford? 終生患癌風(fēng)險(xiǎn):男性攜帶者--67%

GC女性攜帶者---83%

GC女性攜帶者---60%

乳腺小葉癌(LBC)? 18-40ys

攜帶者建議行預(yù)防性全胃切除術(shù)14CDH1與患癌風(fēng)險(xiǎn)(2016NCCN,

V1）? 終生患癌風(fēng)險(xiǎn):男性攜帶者--67%GC14CDH1HDGC中CDH1種系突變的地域差異15? 低風(fēng)險(xiǎn)區(qū)（北美、加拿大、英國）---50%? 中風(fēng)險(xiǎn)區(qū)（德國）---25%? 高風(fēng)險(xiǎn)區(qū)(葡萄牙，意大利)

---22%? 散發(fā)性胃癌高發(fā)區(qū)(中、日、韓)

~10%HDGC中CDH1種系突變的地域差異15? 低風(fēng)險(xiǎn)區(qū)（北美、HDGC中CDH1的突變定位16HDGC中CDH1的突變定位16臨床處理遺傳學(xué)檢測和咨詢

(E-cadherinorCDH1)? 年齡<25歲或早于先證者10年，每6-12個月做一次胃鏡檢查，盲取活檢30塊? 乳腺X光

/乳腺核磁? 預(yù)防性全胃切除vanderPost,etal.Journalof

medicalgenetics,

2015.17臨床處理遺傳學(xué)檢測和咨詢(E-cadherinorCD家族遺傳性胃癌（二）我們的研究18家族遺傳性胃癌（二）我們的研究18Materialand

Methods19Samples:103

HDGC1500sporadic

GC3097normalcontrolMethods:16exonsPCR+

sequencing~~GC：sitesequencing~~Familiesand

controls：MLPA~~HDGCMaterialandMethods19Samples:VariablesTotalNo.ofpatients(N=1603)Age,

years≦3561>35and

<651051≧65491SexMale1145Female458Lauren

classificationIntestinal517Diffuse864GCfamilial

aggregationPositive182Negative1421HDGC(2010

criteria)82HDGC(2015

criteria)9220Results（1）Table1.Patient

informationVariablesTotalNo.ofpatientsc.1018A>G，p.T340A，7

casesc.865G>A p.A289T，1

casec.1273G>C，p.V425L，1case2.CDH1germline

mutationc.1888C>G，

p.L630V，34casesc.2165-1G>A，

1case21c.1018A>G，p.T340A，7casesc.865c.1298A>Gp.D433G,1

casec.2206G>Ap.V736M，

1casec.1103C>GTp.T368S，1

casec.1174G>Ap.V392I，

1casec.1581A>Cp.R527S，

1case22c.1298A>Gp.D433G,1casec.22061case，exon14

deletion3.CDH1rearrangementin

HDGCNormalE14delE14DELNormal23E14del1case，exon14deletion3.CDH1V736MR527SL630VA289TT340AT368SV392IV425LD433G/N4. SitesofCDH1germline

mutations---------------------------------------------------------------------------------------------Missense

mutation2165-1

G>AE14delRearrangement---------------------------------------------------------------------------------------------Splicesite

mutation24V736MR527SL630VA289T4. Siteso5.CDH1L630Vand

GC

25CDH1L630VmutationinGC

andnormal

controlstotalmutation

no.ratePNormal

control3097571.84%GC1591342.14%0.484HDGC（2010）8244.88%0.071HDGC（2015）9144.40%0.0955.CDH1L630VandGC 25CDH1L256.SummaryExonSitesTypePolymorphismFunctional prediction CasesMutationrateMutation

rateinHDGC in

HDGCMutationReferencep.T340Amissense--benign71/82

=0.0121/92

=0.01106253rsnumber MAF PloyPhen SIFT(2010) (2015) ratein

GC-2affect7 c.865G>A

missense - - possibly proteinp.A289T damaging1---- 1/734=0.0014 0function8 c.1018A>G deleteriou 6/1591=0.0s8 c.1103C>GTp.T368Smissensers367868307NAbenigntolerated1---- 1/734=0.00 0149c.1298A>Gp.D433Gmissensers376097289NApossiblyaffectdamaging

proteinfunction1----1/734=0.00140p.V392Imissensers1418640440.0008benigntolerated1----01401406/92=0.06102119 c.1174G>A 1/734=0.0affect9 c.1273G>C

missense - - possibly proteinp.V425L damaging1---- 1/734=0.00function11 c.1581A>Cp.R527S missense - - benign tolerated1---- 1/734=0.0 0014affect12 c.1888C>G probablyp.L630V missense rs2276331 G=0.0012 damaging protein364/82=0.04334/1568=0.functionaffect14 c.2206G>A probably

protein 1 -- -- 1/734=0.00p.V736M missense - - damaging 14

0function14c.2165-1G>Aspliceprobablydamagingaffectproteinfunction11/82=0.141/92=0.111/734=0.0014014E14delrearrangementaffectproteinfunctionaffectproteinfunction11/82=0.141/92=0.11--0266.SummaryExonSitesTypePolymorp26Results（2）CellfunctionstudyforCDH1

L630V27Results（2）27aminoacid

127154708 731

777882EGFR

activationExtracellular

domainJuxtamemberane

domainSrckinase

activationP38

activationL630V28aminoacid 127154708 73177788BFigure1Confirmationofwild-typeormutantCDH1(L630V)FLAGfusionproteinexpressingingastriccancercelllineandCHOcellline(ANCI-N87celllineBCHOcell

line).AFLAGβ-actin(NCI-N87cell

line)FLAGβ-actin(CHOcell

line)29BFigure1Confirmationofwild1.FunctionofL630VinNCI-N87

cell1.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

NCI-N87cell

line.OD

Value

(490nm)00.60.70.824h48h72h96hMockWTL630V0.50.40.30.20.130Figure2CDH1hadnosignificantdifferenceontheproliferationofNCI-N87gastriccancer

cells,Hours1.FunctionofL630VinNCI-N80%20%40%MockWTL630V24h36h

**

** 31Percentage

of

motile

cells1.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

NCI-N87cell

line.Figure3CDH1mutantL630VpromotedthemigrationofNCI-N87gastriccancercellsthroughwoundhealing

assay.A0%20%40%MockWTL630V24h36h *2.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

CHOcell

line.2.FunctionofL630VinCHOcell

line00.80.60.40.211.21.41.624h48h72h96hMockWTL630V32OD

Value

(490nm)Figure4CDH1hadnosignificantdifferenceontheproliferationofCHOcancer

cells,Hours2.1TheinfluenceofCDH1andMockWT2.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

CHOcell

line.0h 12h 24hFigure5CDH1mutantL630VpromotedthemigrationofCHOcancercells

throughwoundhealing

assay.Percentage

of

motile

cells20%L630V0%40%60%MockWTL630V80%12h24h**33MockWT2.2TheinfluenceofCDHDISCUSSIONCDH1anditsmutantL630Vhadno

significantinfluence

on cancercellproliferation

.CDH1mutantL630Vpromotedthemigration

ofgastriccancer

cells.34DISCUSSION34結(jié) 論? 中國人GC中CDH1種系突變以錯義突變?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在我國正常人群和GC、HDGC人群中突變頻率均無顯著差異35結(jié) 論35結(jié) 論36? 90%以上的HDGC

無CDH1種系突變，CDH1種系突變可能不是我國HDGC的主要遺傳易感基因。? 本研究為國內(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)。結(jié) 論36? 90%以上的HDGC無CDH1種系突變，CD家族遺傳性胃癌（三）案例分析37家族遺傳性胃癌（三）案例分析3735號家系

CDH1T340A:carrier:gastriccancerLungcancerEsophegeal

cancerBreastcancerTestedCancerSomaticmutation

T340A61y3834y40y35號家系CDH1T340A:carrier61y38335號家屬，34y，男性，CDH1

T340A3935號家屬，34y，男性，CDH1T340A39688號家系

CDH1L630V40GC:gastric

cancerIGC:intestinal

GCEndC:endometrial

cancer688號家系CDH1L630V40GC:gastric688-241688-241688-442688-442688-843688-843688-944688-944688-1445688-1445建議流

程46? 詳細(xì)地家系咨詢，患者知情同意，遺傳學(xué)檢測? 家系隨訪

“腫瘤易感基因panel”檢測及復(fù)檢? 通知18-40y，無癥狀致病突變攜帶者在遺傳門診咨詢? 建議進(jìn)行胃鏡檢測及取24-30塊活檢、病檢? 根據(jù)結(jié)果進(jìn)一步遺傳門診治療或隨訪建議建議流程46? 詳細(xì)地家系咨詢，患者知情同意，遺傳學(xué)檢4747家族遺傳性胃癌:基因診斷、篩查和臨床處理賈淑芹北京大學(xué)腫瘤醫(yī)院

分子診斷中心1家族遺傳性胃癌:基因診斷、篩查和臨床處理賈淑芹48AutosomalDominantInheritedCancer

Syndromes?Breast

and

Ovarian

Cancer

BRCA1&2ColonCancerand

Polyposis49HNPCCFAPPolyposisCowdensPeutz-JehgersJuvenile

PolyposisMMRAPCMYHPTENSTK11SMAD4BMPR1AOtherGI

CancersGastricPancreasMEN1MEN2/MTCVHLLi-FraumeniCDH1p16MeninRETVHLp53北京大學(xué)腫瘤醫(yī)院（PKUCH)分子診斷中心(MDC)癌癥遺傳基因篩查101基因panel為癌癥患者及其家系成員進(jìn)行風(fēng)險(xiǎn)評估、遺傳咨詢和干預(yù)AutosomalDominantInherited4950350癌癥的遺傳易感性51? 誰應(yīng)該做癌癥遺傳易感性檢測？? 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？? 對于未發(fā)病的攜帶者，我們能做些什么？癌癥的遺傳易感性4? 誰應(yīng)該做癌癥遺傳易感性檢測？? 若攜帶51?遺傳彌漫型胃癌(HDGC)?林奇綜合征(Lynch)?遺傳性乳腺癌卵巢癌綜合征(HBOC)?青少年息肉綜合征(JPS

)?黑斑息肉綜合征(

PJS)90%?家族性腺瘤息肉病(FAP

)?李-佛美尼綜合征（LFS)?……家族遺傳性胃癌（-）背景52?遺傳彌漫型胃癌(HDGC)家族遺傳性胃癌（-）背景552家族性胃癌的遺傳風(fēng)險(xiǎn)53Chun&Ford,CancerJ.

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%家族性胃癌的遺傳風(fēng)險(xiǎn)6Chun&Ford,Cancer53遺傳性彌漫型胃癌? 1998年，首次發(fā)現(xiàn)于新西蘭3個毛利家族中，早發(fā)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16q22.1? 在3個家系中都發(fā)現(xiàn)E-cadherin

(CDH-1）種系截短突變? 外顯率：60歲前，70%的患者均患癌54遺傳性彌漫型胃癌? 1998年，首次發(fā)現(xiàn)于新西蘭3個毛利家族54遺傳性彌漫型胃癌

常染色體顯性遺傳

早發(fā):

14~69y, 平均:

37yLauren分型:

彌散型

不易早期診斷，預(yù)后差? CDH1

種系突變?yōu)樘卣?5遺傳性彌漫型胃癌 常染色體顯性遺傳855HDGC

診斷標(biāo)準(zhǔn)（IGCLC)2.3confirmedDGCcasesin1stor2nddegreerelativesindependentofageSingle

DGC<40PersonalorfamilyhistoryofDGCandLBC,1

case<50Single

DGC<40PersonalorfamilyhistoryofDGCandLBC,1case<5020101. 2GCcases

in

family,oneconfirmed

DGC<5020151.2GCcasesinfamily,oneconfirmed

DGC19991.2GCcasesinfamily,oneconfirmed

DGC<502.3confirmedDGCcasesin1stor2nddegreerelativesindependentofage56HDGC診斷標(biāo)準(zhǔn)（IGCLC)2.3confirmed562015

版標(biāo)準(zhǔn)解讀57? 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? 任何一個家屬同時具有彌漫型胃癌和乳腺小葉癌，其中至少一例<50歲? 個人雙側(cè)乳腺小葉癌或者多個家族成員乳腺小葉癌，一例<

50歲? 患者同時具有彌漫型胃癌和唇/腭裂? 印戒細(xì)胞癌的癌前病變2015版標(biāo)準(zhǔn)解讀10? 2例以上胃癌，至少其中一例是彌漫575811CDH1突變的患者印戒細(xì)胞癌中E-cadherin表達(dá)降低或缺如59CDH1突變的患者印戒細(xì)胞癌中E-cadherin表12CDH1

與患癌風(fēng)險(xiǎn)? 終生患癌風(fēng)險(xiǎn):男性攜帶者--70%

GC女性攜帶者---56%

GC女性攜帶者---42%

乳腺小葉癌(LBC)? 中位發(fā)病年齡

毛利族:32

yrs其他：

43yrs60Hansford&Huntsman,JAMAOnc

2015CDH1與患癌風(fēng)險(xiǎn)? 終生患癌風(fēng)險(xiǎn):13Hansford? 終生患癌風(fēng)險(xiǎn):男性攜帶者--67%

GC女性攜帶者---83%

GC女性攜帶者---60%

乳腺小葉癌(LBC)? 18-40ys

攜帶者建議行預(yù)防性全胃切除術(shù)61CDH1與患癌風(fēng)險(xiǎn)(2016NCCN,

V1）? 終生患癌風(fēng)險(xiǎn):男性攜帶者--67%GC14CDH1HDGC中CDH1種系突變的地域差異62? 低風(fēng)險(xiǎn)區(qū)（北美、加拿大、英國）---50%? 中風(fēng)險(xiǎn)區(qū)（德國）---25%? 高風(fēng)險(xiǎn)區(qū)(葡萄牙，意大利)

---22%? 散發(fā)性胃癌高發(fā)區(qū)(中、日、韓)

~10%HDGC中CDH1種系突變的地域差異15? 低風(fēng)險(xiǎn)區(qū)（北美、HDGC中CDH1的突變定位63HDGC中CDH1的突變定位16臨床處理遺傳學(xué)檢測和咨詢

(E-cadherinorCDH1)? 年齡<25歲或早于先證者10年，每6-12個月做一次胃鏡檢查，盲取活檢30塊? 乳腺X光

/乳腺核磁? 預(yù)防性全胃切除vanderPost,etal.Journalof

medicalgenetics,

2015.64臨床處理遺傳學(xué)檢測和咨詢(E-cadherinorCD家族遺傳性胃癌（二）我們的研究65家族遺傳性胃癌（二）我們的研究18Materialand

Methods66Samples:103

HDGC1500sporadic

GC3097normalcontrolMethods:16exonsPCR+

sequencing~~GC：sitesequencing~~Familiesand

controls：MLPA~~HDGCMaterialandMethods19Samples:VariablesTotalNo.ofpatients(N=1603)Age,

years≦3561>35and

<651051≧65491SexMale1145Female458Lauren

classificationIntestinal517Diffuse864GCfamilial

aggregationPositive182Negative1421HDGC(2010

criteria)82HDGC(2015

criteria)9267Results（1）Table1.Patient

informationVariablesTotalNo.ofpatientsc.1018A>G，p.T340A，7

casesc.865G>A p.A289T，1

casec.1273G>C，p.V425L，1case2.CDH1germline

mutationc.1888C>G，

p.L630V，34casesc.2165-1G>A，

1case68c.1018A>G，p.T340A，7casesc.865c.1298A>Gp.D433G,1

casec.2206G>Ap.V736M，

1casec.1103C>GTp.T368S，1

casec.1174G>Ap.V392I，

1casec.1581A>Cp.R527S，

1case69c.1298A>Gp.D433G,1casec.22061case，exon14

deletion3.CDH1rearrangementin

HDGCNormalE14delE14DELNormal70E14del1case，exon14deletion3.CDH1V736MR527SL630VA289TT340AT368SV392IV425LD433G/N4. SitesofCDH1germline

mutations---------------------------------------------------------------------------------------------Missense

mutation2165-1

G>AE14delRearrangement---------------------------------------------------------------------------------------------Splicesite

mutation71V736MR527SL630VA289T4. Siteso5.CDH1L630Vand

GC

72CDH1L630VmutationinGC

andnormal

controlstotalmutation

no.ratePNormal

control3097571.84%GC1591342.14%0.484HDGC（2010）8244.88%0.071HDGC（2015）9144.40%0.0955.CDH1L630VandGC 25CDH1L726.SummaryExonSitesTypePolymorphismFunctional prediction CasesMutationrateMutation

rateinHDGC in

HDGCMutationReferencep.T340Amissense--benign71/82

=0.0121/92

=0.01106253rsnumber MAF PloyPhen SIFT(2010) (2015) ratein

GC-2affect7 c.865G>A

missense - - possibly proteinp.A289T damaging1---- 1/734=0.0014 0function8 c.1018A>G deleteriou 6/1591=0.0s8 c.1103C>GTp.T368Smissensers367868307NAbenigntolerated1---- 1/734=0.00 0149c.1298A>Gp.D433Gmissensers376097289NApossiblyaffectdamaging

proteinfunction1----1/734=0.00140p.V392Imissensers1418640440.0008benigntolerated1----01401406/92=0.06102119 c.1174G>A 1/734=0.0affect9 c.1273G>C

missense - - possibly proteinp.V425L damaging1---- 1/734=0.00function11 c.1581A>Cp.R527S missense - - benign tolerated1---- 1/734=0.0 0014affect12 c.1888C>G probablyp.L630V missense rs2276331 G=0.0012 damaging protein364/82=0.04334/1568=0.functionaffect14 c.2206G>A probably

protein 1 -- -- 1/734=0.00p.V736M missense - - damaging 14

0function14c.2165-1G>Aspliceprobablydamagingaffectproteinfunction11/82=0.141/92=0.111/734=0.0014014E14delrearrangementaffectproteinfunctionaffectproteinfunction11/82=0.141/92=0.11--0266.SummaryExonSitesTypePolymorp73Results（2）CellfunctionstudyforCDH1

L630V74Results（2）27aminoacid

127154708 731

777882EGFR

activationExtracellular

domainJuxtamemberane

domainSrckinase

activationP38

activationL630V75aminoacid 127154708 73177788BFigure1Confirmationofwild-typeormutantCDH1(L630V)FLAGfusionproteinexpressingingastriccancercelllineandCHOcellline(ANCI-N87celllineBCHOcell

line).AFLAGβ-actin(NCI-N87cell

line)FLAGβ-actin(CHOcell

line)76BFigure1Confirmationofwild1.FunctionofL630VinNCI-N87

cell1.1TheinfluenceofCDH1anditsmutantL630Vontheproliferationin

NCI-N87cell

line.OD

Value

(490nm)00.60.70.824h48h72h96hMockWTL630V0.50.40.30.20.177Figure2CDH1hadnosignificantdifferenceontheproliferationofNCI-N87gastriccancer

cells,Hours1.FunctionofL630VinNCI-N80%20%40%MockWTL630V24h36h

**

** 78Percentage

of

motile

cells1.2TheinfluenceofCDH1anditsmutantL630Vonthemigrationin

NCI-N87cell

line.Figure3CDH1mutantL630VpromotedthemigrationofNCI-N87gastriccancercellsthroughwoundhealing

assay.A0%20%40%MockWTL630V24h36h *2.1TheinfluenceofCDH1andits