现代消化及介入诊疗
現代消化及介入診療
현대소화급개입진료
MODERN DIGESTION & INTERVENTION
2013年
4期
205-207
,共3页
薛净%沙卫红%聂玉强%李瑜元
薛淨%沙衛紅%聶玉彊%李瑜元
설정%사위홍%섭옥강%리유원
siRNA%Survivin基因%HepG2细胞%细胞凋亡
siRNA%Survivin基因%HepG2細胞%細胞凋亡
siRNA%Survivin기인%HepG2세포%세포조망
siRNA%Survivin%HepG2 cells%Apoptosis
目的观察特异小干扰RNA(small interfering RNA, siRNA)抑制survivin基因表达对人肝癌细胞株HepG2细胞凋亡的影响。方法构建survivin-siRNA真核表达载体,利用脂质体转染人肝癌HepG2细胞,采用反转录聚合酶链反应(RT-PCR)技术检测survivin基因mRNA表达水平。针对转染后不同的时间点采用四甲基偶氮唑蓝(MTT)法检测各组细胞的增殖以及HepG2细胞的凋亡情况。结果构建的survivin-siRNA转染组survivin蛋白表达较对照组明显下调,差异有显著性(P<0.01),其增殖能力较对照组显著下降(P<0.001);而阳性对照组及未转染组对survivin表达及mRNA抑制作用不明显(P=0.219)。结论所构建的针对人survivin-siRNA可显著抑制survivin的表达及HepG2细胞凋亡。
目的觀察特異小榦擾RNA(small interfering RNA, siRNA)抑製survivin基因錶達對人肝癌細胞株HepG2細胞凋亡的影響。方法構建survivin-siRNA真覈錶達載體,利用脂質體轉染人肝癌HepG2細胞,採用反轉錄聚閤酶鏈反應(RT-PCR)技術檢測survivin基因mRNA錶達水平。針對轉染後不同的時間點採用四甲基偶氮唑藍(MTT)法檢測各組細胞的增殖以及HepG2細胞的凋亡情況。結果構建的survivin-siRNA轉染組survivin蛋白錶達較對照組明顯下調,差異有顯著性(P<0.01),其增殖能力較對照組顯著下降(P<0.001);而暘性對照組及未轉染組對survivin錶達及mRNA抑製作用不明顯(P=0.219)。結論所構建的針對人survivin-siRNA可顯著抑製survivin的錶達及HepG2細胞凋亡。
목적관찰특이소간우RNA(small interfering RNA, siRNA)억제survivin기인표체대인간암세포주HepG2세포조망적영향。방법구건survivin-siRNA진핵표체재체,이용지질체전염인간암HepG2세포,채용반전록취합매련반응(RT-PCR)기술검측survivin기인mRNA표체수평。침대전염후불동적시간점채용사갑기우담서람(MTT)법검측각조세포적증식이급HepG2세포적조망정황。결과구건적survivin-siRNA전염조survivin단백표체교대조조명현하조,차이유현저성(P<0.01),기증식능력교대조조현저하강(P<0.001);이양성대조조급미전염조대survivin표체급mRNA억제작용불명현(P=0.219)。결론소구건적침대인survivin-siRNA가현저억제survivin적표체급HepG2세포조망。
Objective To study the effect of small interfering RNA (siRNA)of human survivin gene on apoptosis of human hepatoma cell line HepG2. Methods The PCR products containing siRNA sequence were transfected into HepG2 via Lipofectamine TM 2000, cell viability was determined by MTT assay. Apoptosis was detected by morphological observation and flow cytometry analysis. The expression level of HepG2 mR-NA was assayed by RT-PCR. Result The cell growth and viability of siRNA transfected group were signifi-cantly inhibited when compared with that of the negative and blank control groups(P<0.01), whereas the latter two groups had simi1ar expression levels. Conclusion Interference with siRNA against survivin gene can ef-fectively inhibit the specific gene expression, which may play a significant role in inducing of apoptosis of the corresponding HepG2 cells.