livin基因沉默对大肠癌细胞凋亡和化疗敏感性的影响

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1、Livin基因沉默对大肠癌细胞凋亡和化疗敏感性的影响作者：严红彭淑梅罗其泰邱春雷延卫东【摘要】目的观察沉默Livin基因对大肠癌HT-29细胞凋亡和化疗敏感性的影响。方法采用脂质体转染技术将化学合成的LivinsiRNA转入HT-29细胞，RT-PCR和Western印迹法检测转染效果。选用5-氟尿嘧啶(5-FU)作用于转染和未转染的细胞，MTT检测各组细胞的增殖、流式细胞仪检测细胞的凋亡。结果si-Livin1转染组LivinαmRNA、LivinβmRNA和Livin蛋白表达水平较其余干扰组和对照组显著降低(P<0.01

2、)。5-FU对HT-29细胞的生长抑制作用强于si-Livin1转染组(P<0.01)，而si-Li　　vin1+5-FU组对细胞的生长抑制率显著高于单独si-Livin1转染组和单独5-FU作用组(P<0.01)。与空白对照组比较，si-Livin1转染可使细胞出现明显凋亡现象(P<0.01)，5-FU对细胞的促凋亡作用强于si-Livin1转染组(P<0.01)，si-Livin1+5-FU组的凋亡率显著高于单独si-Livin1转染组和单独5-FU作用组(P<0.01)。结论siRNA沉默Liv

3、in基因能够抑制HT-29细胞的生长，促其凋亡，提高5-FU的化疗敏感性。Livin基因有望成为大肠癌治疗的新靶点。【关键词】Livin;大肠癌;凋亡抑制蛋白;RNA干扰;细胞凋亡;化疗敏感性13　【Abstract】ObjectiveToexploretheeffectsofLivingenesilencingonchemotherapeuticsensitivityandapoptosisofcolorectalcarcinomaHT-29cells.MethodsLivinspecificsiRNAoligonucleoti

4、desweredesignedandsynthesizedartificially.SiRNAwastransfectedintocolorectalcarcinomaHT-29cellsbylipofectiontechnology.TransfectionefficiencywasdetectedbyWesternblotandRT-PCR.Thetransfectedanduntransfectedcellswereincubatedinthepresenceof5-FU.Cellularproliferationactiv

5、etieswereassayedbyMTT.Theapoptosisofcellswasassayedbyflowcytometry(FCM).ResultsComparedwiththoseofothertransfectedg　roupsandcontrolgroup,theproteinlevelsofLivinandthemRNAexpressionsofLivinαandLivinβweredecreasedsignificantlyaftertransfectedbysi-Livin1(P<0.01).Thegr

6、owthinhibitionratesofcellsincubatedin5-FUwerehigherthanthoseoftransfectedbysi-Livin1(P<0.01).Thegrowthinhibitionratesofcellsaffectedbysi-Livin1and5-FUwerehigherthanthoseofsingletransfectedbysi-Livin1andsingleincubatedin5-FUsignificantly(P<0.01).Theapoptosisrates

7、ofcellstransfectedbysi-Livin1werehigherthanthoseofcontrolgroup(P<0.01),butlowerthanthoseofincubatedin135-FU(P<0.01).Theapoptosisratesofcellsaffectedbysi-Livin1and5-FUwerehigherthanthoseofsingletransfectedbysi-Livin1andsingleincubatedin5-FUsignificantly(P<0.01

8、).　ConclusionsLivingenesilencedbysiRNAinducesgrowthsuppressionandapoptosisofHT-29cells,whichcouldincreasethechemotherapeutic