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《靶向fak的sirna對喉癌細(xì)胞增殖及侵襲能力的抑制作用》由會員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫。
1、靶向FAK的siRNA對喉癌細(xì)胞增殖及侵襲能力的抑制作用張潛英，葉琳，余曉燕，沈娜，鄧碧，吳曉松，陳鴻雁（重慶醫(yī)科大學(xué)附屬第一醫(yī)院耳鼻咽喉科,重慶400016）摘要：目的運(yùn)用RNA干擾(RNAinterference，RNAi)技術(shù)阻斷喉癌Hep-2細(xì)胞株中FAK基因的表達(dá)，并研究FAK基因沉默后對Hep-2細(xì)胞株產(chǎn)生的影響。方法針對FAK基因，構(gòu)建2條干擾重組質(zhì)粒FAK-pGenesil-CH1和FAK-pGenesil-CH2，1條陰性對照重組質(zhì)粒FAK-pGenesil-HK；在脂質(zhì)體的介導(dǎo)下轉(zhuǎn)染喉癌He
2、p-2細(xì)胞株，用RT-PCR和Westernblot分析FAKmRNA及蛋白的表達(dá)；MTT法、流式細(xì)胞技術(shù)及體外侵襲實(shí)驗(yàn)測定干擾重組質(zhì)粒對Hep-2細(xì)胞株的增殖、侵襲能力的影響。結(jié)果重組干擾質(zhì)粒能有效地阻斷Hep-2細(xì)胞株中FAK基因在mRNA和蛋白水平上的表達(dá)(P＜0.05)；重組質(zhì)粒轉(zhuǎn)染Hep-2細(xì)胞后，細(xì)胞增殖抑制率分別為58.34%、52.78%，與對照組比較差別有統(tǒng)計(jì)學(xué)意義（P＜0.05）；干擾組G0～G1期細(xì)胞增多，S期細(xì)胞數(shù)量明顯減少（P＜0.05）；FAK-pGenesil-CH1、FAK-pG
3、enesil-CH2作用Hep-2細(xì)胞株48h后，Hep-2細(xì)胞穿過Matrigel膜的個數(shù)分別為（56.0±6.7）、（51.0±5.1），與陰性對照組及正常對照組[分別為（152.0±9.4）和（139.0±8.1）]比較有統(tǒng)計(jì)學(xué)差別（P＜0.05）。結(jié)論干擾重組質(zhì)粒能有效地抑制轉(zhuǎn)染Hep-2細(xì)胞株的FAKmRNA及蛋白的表達(dá)，并能抑制Hep-2細(xì)胞株的增殖及侵襲能力。關(guān)鍵詞：FAK；siRNA；RNA干擾；Hep-2細(xì)胞株；增殖；侵襲中圖法分類號：R739.65文獻(xiàn)標(biāo)識碼：AEffectofFAKgene
4、silencingonproliferationandinvasioninofHep-2cellsZHANGQian-ying,YELin,YUXiao-yan,SHENNa,DENGBi,WUXiao-song,CHENHong-yan(DepartmentofOtolaryngology,theTheFirstAffilatedHospital，,ChongqingMedicalUniversity，,Chongqing400016，,China)Abstract:objectiveObjectiveToo
5、bservetheeffectsofRNAi-mediatedFAKgeneonsilencinginonlaryngealcarcinomaHep-2cells．.MethodsConstructrRecombinantplasmidsgeneratingshorthairpinRNAwereconstructed,includingtwointerferentialsequencesFAK-pGenesil-CH1、,FAK-pGenesil-CH2andonenegativecontrolsequence
6、FAK-pGenesil-HK.Hep-2cellsweretransfectedwithrecombinantplasmids．.FAKmRNAandFAKproteinwereanalyzedrespectivelybyRT-PCRandWesternblotting,respectively．.Cellproliferation、,cellcycleandinvasionofHep-2cellswereobservedbyMTTassay、,flowcytometry（FCM）andtranswellin
7、vasionassay,respectively.ResultsRecombinantplasmidsofFAK-pGenesil-CH1andFAK-pGenesil-CH2downregulatedtheexpressionsofFAKmRNAandprotein(P<0.05)；.TheproliferationinhibitionratioinHep-2cellsreachedto58.34%、and52.78%,%48hourshaftertransfectionofrecombinantplasmi
8、dsofFAKpGenesil-CH1、andFAK-pGenesil-CH2,whilethatthecontrolwas8.34%inthecontrol(P<0.05)（P<0.05）.;FCManalysisshowedthatmMorecellsstayarrestedatinG0-/G1phaseinthetransfectedgroupsthancontrols;.The
