黏着斑激酶通过细的胞周期蛋白依赖性激酶2、cjunn端激酶促进人肺动脉平滑肌细胞增殖.doc

黏着斑激酶通过细胞周期蛋白依赖性激酶2、c-junN端激酶促进人肺动脉平滑肌细胞增殖 中国病理生理杂志ChineseJournalofPathophysiology2008,24(2):303—306 [文章编号]1000—4718(2008)02—0303—04 黏着斑激酶通过细胞周期蛋白依赖性激酶2, C-junN端激酶促进人肺动脉平滑肌细胞增殖 林春龙 (湖南师范大学附属岳阳医院,岳阳市立二医院呼吸内科,湖南岳阳414000) ? 303? [摘要]目的:探讨黏着斑激酶(FAK)促进人肺动脉平滑肌细胞(HPASMCs)增殖的机制.方法:取肺癌 部分切除手术病人远离肺癌的周围正常组织,酶消化法培养细胞.在正义寡核苷酸(FAKSODNs),反义寡核苷酸 (FAKASODNs),随机寡核苷酸(FAKMS)转染后,用免疫印迹方法测定了FAK,C—junN端激酶(JNK),细胞周期 蛋白依赖性激酶2(CDK2)的表达;用流式细胞仪测定了细胞周期,细胞凋亡;用免疫组织化学法测定胞浆FAK的 表达.结果:免疫印迹结果表明,与FAKMS组比较,应用FAKASODNs转染后HPASMCsFAK,JNK,CDK2蛋白质 的表达明显降低(Plt;0.01,Plt;0.05,Plt;0.01);而用正义寡核苷酸转染后则明显增加(Plt;0.01,Plt;0.05, Plt;0.05).流式细胞仪测定结果示:与FAKMS转染组比较:FAKSODNs转染后,HPASMCs中G.相的比例显着 减少,S相显着增加(Plt;0.01),细胞凋亡减少(Plt;0.01);而FAKASODNs转染后结果则相反.免疫组织化学结 果显示:FAKSODNs转染后HPASMCs胞浆染色增强,FAKASODNs转染后胞浆浅染,提示FAK表达减少.结论: FAK可能经由JNK,CDK2信号通路促使细胞增殖. [关键词]黏着斑激酶;人肺动脉平滑肌细胞;C—junN端激酶;细胞周期蛋白依赖激酶类 [中图分类号]R363[文献标识码]A CDKandJNKmediateFAKtoenhanceproliferationinhumanpuimona- ryarterysmoothcells LINChun—long (RespiratoryDepartment,theSecondHospitalofYueyangCity,theMedwalCollegeofHunanNormalUniversity,Yueyang 414000,China) [ABSTRACT]AIM:Tofindoutthemechanismoffocaladhesionkinase(FAK)facilitatinghumanpulmonaryar— terysmoothmusclecells(HPASMCs)proliferation.METHODS:HPASMCswereisolatedfromnormalpartoflungsoftwo carcinomapatientswhoundergonelungpartialresection.CulturedHPASMCsstimulatedbyfibronection(40mg/L)were passivelytransfectedwithODNs,sensefocaladhesionkinase(FAK),mismatchsenseandantisense—FAK,respectively. ExpressionofFAK,JunNH2一terminalkinase(JNK)andcyclin—dependentkinase2(CDK2)proteinsweredetectedby immunoprecipitationandWesternblotting.Cellcycleandcellapoptosiswereanalyzedbyflowcytometry.Inaddition,cyto— plasmaFAKexpressionwasdetectedbyimmunohistochemistrystaining.RESULTS:TheproteinexpressionsofFAK,JNK andCDK2inHPASMCsdecreasedinFAKASODNsgroupandincreasedinFAKSODNsgroup.Meanwhile,theproportion ofcellsatG1phasedecreasedsignificantlyinFAKSODNsgroup,whilethecellsatSphaseincreasedsignificantly.Incon- trast,theproportionofcellsatG1phaseWas