GPC-3特异性短发夹型RNA干预基因转录对肝癌细胞增殖与凋亡的影响-论文.pdf

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1、实用肝脏病杂志2013年10月第l6卷第5期JClinHepatol，Oct．2013．Vo1．16No．5·401··肝肿瘤·GPC一3特异性短发夹型RNA干预基因转录对肝癌细胞增殖与凋亡的影响邰伯军姚敏蔚丹丹陈洁时运顾星张洁邱历伟姚登福【摘要】目的观察磷脂酰肌醇蛋白多糖(GPC)一3基因特异性短发夹型RNA(shRNA)对HepG2细胞增殖的抑制作用。方法设计、合成和筛选GPC一3特异的高抑制率shRNA并构建质粒；观察shRNA沉默GPC一3基因对肝癌细胞增殖、细胞周期及凋亡的影响。结果成功构建了4种GPC一3shRNA

2、干扰质粒，以脂质体介导分别转染HepG2细胞后，筛选出的shRNA干扰质粒抑制率最高达89．3％；以该高效GPC一3shRNA质粒转染HepG2细胞24h、48h和72h后，细胞增殖较对照组分别下降了33．2％、56．0％和71．1％(尸<0．05)，细胞多被阻滞在G1期；HepG2细胞凋亡率达65．6％，显著高于对照组(约为5％，P<0．05o结论GPC一3特异性的shRNA可有效降低HepG2细胞中GPC一3基因转录水平，抑制增殖并促进其凋亡。因此，GPC一3可望成为肝癌基因治疗的靶目标。【关键词】HepG2细胞；磷脂酰肌

3、醇蛋白多糖一3；短发夹型RNA；基因沉默doi：10．3969~．issn．1672-5069．2013．05．006Efectsofglypican——3geneknockdownbyshRNAonproliferationandapoptosisofHepG2cellsTa／Bojun，YaoMin，YuDandan，eta1．ResearchCenterofClinicalMedicine，AfiliatedHospitaltONantongUniversity，Nantong226001，JiangsuProvince

4、，China【Abstract】ObjecfiveToinvestigatetheeffectsofglypican-3(GPC-3)geneknockdownbyspecificshorthairpinRNA(shRNA)onproliferationandapoptosisofHepG2cells．MethodsFourpairsofGPC-3shRNAweredesigned．synthesizedandscreenedaccordingtotheinhibitingefficiencyandplasmidscontai

5、ningspecificGPC一3shRNAwereconstructed；HepG2cellsweretransfectedwithconstructedplasmidcontainingGPC一3shRNAwiththehighestinhibitingrate，andcellproliferation，cellcycletransition，andapoptosisweredeterminedafterGPC一3knockdown．ResultsFourplasmidscontainingspecificGPC一3shR

6、NAweresuccessfullyconstructed，andoneofwhichhadthehighestinhibitingratebyreachingto89-3％；TheproliferationratesoftheHepG2cellstransfectedwiththeeficientGPC一3shRNAdecreasedto33．2％，56．O％and71．1％，respectively，ascomparedtothecore—spondingcontrols(P<0．05)24h，48hand72hoursa

7、ftertransfection；ThecellcycleswerearrestedatG1phaseaf-terGPC一3knockdown．whiletheapoptosisrateoftransfectedHepG2cellsincreasedto65．6％，significantlyhigherthanthatincontrol(5％．P

8、s，resultingincellproliferationinhibition，cellcyclearestandincreasedapoptosis；GPC一3mightbeapotentialtargetgeneforhepatocellularcarcinomathe