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1、接合物蛋白CrkⅠ在卵巢癌恶性潜能中的作用车亚玲,王瑾,令狐华(400016重庆,重庆医科大学附属第一医院妇产科)[摘要]目的证实接合物蛋白CrkⅠ在卵巢癌恶性潜能中的作用。方法采用Westernblot法检测卵巢癌组织、卵巢良性肿瘤组织、正常卵巢组织中Crk和Dock180蛋白的表达；用免疫沉淀法检测3种卵巢癌细胞株中Crk与Dock180蛋白的内源性结合；用小干扰RNA敲低SKOV3细胞中内源性的Crk，检测各组细胞Crk蛋白表达水平、Rac1酶活性和侵袭力的变化。结果Dock180与CrkⅠ的表达强度呈现明显的一致性，卵巢癌组织中二者的表达均显
2、著高于卵巢良性肿瘤组织和正常卵巢组织；而在卵巢良性肿瘤组织与正常卵巢组织之间未发现明显差异。3个卵巢癌细胞株中Dock180主要表现出与CrkⅠ的内源性结合。和对照相比，敲低Crk基因的细胞表现出侵袭能力和Rac1酶活性明显下降(P<0.05)，而敲低CrkⅠ的细胞和同时敲低CrkⅠ和CrkⅡ的细胞相比无明显差异(P>0.05)。结论卵巢癌的恶性生物学行为与Crk/Dock180/Rac1信号通路相关，其中CrkⅠ在卵巢癌中扮演着主要角色。[关键词]信号接合物蛋白Crk；Dock180；人卵巢癌[中图法分类号][文献标志码]AAdaptorprote
3、inCrkⅠmediatesthemalignantpotentialofhumanovariancancerCheYaling,WangJin,LinghuHua(DepartmentofObstetricsandGynecology,theFirstAffiliatedHospitalofChongqingMedicalUniversity,Chongqing,400016,China)[Abstract]ObjectiveToverifytheroleofCrkⅠinmalignantpotentialofhumanovariancancer.
4、MethodsCrkandDock180expressionweredetectedbyWesternblottinginovariancancertissues(EOC,n=28),benignovariantumors(BOT,n=13)andnormalovarytissues(Normal,n=10).Co-precipitationwasperformedtoevaluatetheinvivoprotein-proteininteractionofDock180andCrkinthreeovariancancercells.Theexpre
5、ssionofCrkinSKOV3cellswereknockdownbyusingsiRNA,andtheirRac1activityandthuscellinvasionwereobserved.ResultsTheintensityofCrkⅠandDock180expressionwasconsistentwitheachotherinEOCtissues.BothwereobservedtobesignificantlyhigherinEOCthanthoseintheBOTandnormalovarytissues.Nosignifica
6、ntdifferencewasfoundbetweenBOTandNormalgroupeitherforCrkⅠorDock180expression.Inconsistentwiththisresult,Dock180preferredtocombinewithCrkⅠratherthanwithCrkⅡinallthreeovariancancercells.Furthermore,CrkknockdowncellspresentedwithsustainableCrkⅠexpressiondepletion,significantlydecr
7、easedRac1activityandcellinvasion.ConclusionCrkmightbeinvolvedinmalignantpotentialofhumanEOCmainlythroughCrkⅠ/Dock180/Rac1pathway.[Keywords]adaptorproteinCrkⅠ;CrkⅡ;Dock180;Rac1;humanovariancancerSupportedbytheGeneralProgramofNationalNaturalScienceFoundationofChina(30672432;30772
8、330)andtheFundfromtheFirstAffiliatedHospitalofChongqin
