中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2009年
1期
47-49
,共3页
廖允军%王成友%张敏杰%倪勇%程书榜%詹勇强%范子冰
廖允軍%王成友%張敏傑%倪勇%程書榜%詹勇彊%範子冰
료윤군%왕성우%장민걸%예용%정서방%첨용강%범자빙
CDC20%RNA干扰%癌,肝细胞%有丝分裂
CDC20%RNA榦擾%癌,肝細胞%有絲分裂
CDC20%RNA간우%암,간세포%유사분렬
CDC20%RNA interference%Carcinoma,hepatoeellular%Mitosis
目的 观察RNA干扰(RNAi)对肝癌细胞株Hepal-6中CDC20表达的抑制作用.方法 设计针对小鼠肝癌细胞Hepal-6 CDC20 mRNA的小干扰RNA(siRNA),以阳离子脂质体包埋后转染鼠肝癌细胞Hepal-6,转染48 h后,采用逆转录-聚合酶链反应(RT-PCR)和Western blot方法 检测Hepal-6细胞CDC20 mRNA及蛋白质表达的变化.结果 siRNA各组与正常对照组和阴性对照组比较,CDC20 mRNA的表达均受明显抑制,并呈剂量依赖性(在100、50、25 nmol/L浓度下,Mixed siRNA组与正常对照组mRNA表达比较:0.370±0.058比0.840±0.044,0.480±0.081比0.900±0.068,0.830±0.062比1.010±0.054,各组P＜0.01);CDC20蛋白质的表达亦均受抑制(siRNA-1、siRNA-2、siRNA-3及Mixed组与N-Cont或Blank组蛋白质表达比较:0.482±0.035/0.402±0.003/0.479±0.026/0.446±0.017比0.826±0.03l或0.835±0.024,各组P均＜0.01).结论 CDC20 siRNA可显著抑制小鼠肝癌细胞Hepal-6中CDC20 mRNA和蛋白质的表达.
目的 觀察RNA榦擾(RNAi)對肝癌細胞株Hepal-6中CDC20錶達的抑製作用.方法 設計針對小鼠肝癌細胞Hepal-6 CDC20 mRNA的小榦擾RNA(siRNA),以暘離子脂質體包埋後轉染鼠肝癌細胞Hepal-6,轉染48 h後,採用逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot方法 檢測Hepal-6細胞CDC20 mRNA及蛋白質錶達的變化.結果 siRNA各組與正常對照組和陰性對照組比較,CDC20 mRNA的錶達均受明顯抑製,併呈劑量依賴性(在100、50、25 nmol/L濃度下,Mixed siRNA組與正常對照組mRNA錶達比較:0.370±0.058比0.840±0.044,0.480±0.081比0.900±0.068,0.830±0.062比1.010±0.054,各組P＜0.01);CDC20蛋白質的錶達亦均受抑製(siRNA-1、siRNA-2、siRNA-3及Mixed組與N-Cont或Blank組蛋白質錶達比較:0.482±0.035/0.402±0.003/0.479±0.026/0.446±0.017比0.826±0.03l或0.835±0.024,各組P均＜0.01).結論 CDC20 siRNA可顯著抑製小鼠肝癌細胞Hepal-6中CDC20 mRNA和蛋白質的錶達.
목적 관찰RNA간우(RNAi)대간암세포주Hepal-6중CDC20표체적억제작용.방법 설계침대소서간암세포Hepal-6 CDC20 mRNA적소간우RNA(siRNA),이양리자지질체포매후전염서간암세포Hepal-6,전염48 h후,채용역전록-취합매련반응(RT-PCR)화Western blot방법 검측Hepal-6세포CDC20 mRNA급단백질표체적변화.결과 siRNA각조여정상대조조화음성대조조비교,CDC20 mRNA적표체균수명현억제,병정제량의뢰성(재100、50、25 nmol/L농도하,Mixed siRNA조여정상대조조mRNA표체비교:0.370±0.058비0.840±0.044,0.480±0.081비0.900±0.068,0.830±0.062비1.010±0.054,각조P＜0.01);CDC20단백질적표체역균수억제(siRNA-1、siRNA-2、siRNA-3급Mixed조여N-Cont혹Blank조단백질표체비교:0.482±0.035/0.402±0.003/0.479±0.026/0.446±0.017비0.826±0.03l혹0.835±0.024,각조P균＜0.01).결론 CDC20 siRNA가현저억제소서간암세포Hepal-6중CDC20 mRNA화단백질적표체.
Objective To evaluate the inhibitory effect of CDC20 siRNA on the expression of CDC20 mRNA and protein in mouse Hepal-6 hepatocarcinoma cell lines.Methods The siRNAs which aimed at CDC20 mRNA of Hepal-6,were designed and synthesized.Using cation lipofectamine 2000 embedding techniques,the siRNAs were transfected into Hepal-6 cell lines for 48 h,and then the expression of CDC20 mRNA and protein was detected by RT-PCR and Western blot.Results Compared with normal and negative control groups,CDC20 mRNA expression in the siRNA group was obviously decreased with the increased concentrations of siRNAs.Under the concentrations of 100,50 and 25 nmol/L siRNA,mRNA expression levels in mixed siRNA group and normal control group were 0.370±0.058 vs 0.840±0.044,0.480±0.081 vs 0.900±0.068,and 0.830±0.062 vs 1.010±0.054 respectively(P＜0.01).CDC20 protein expression levels in the siRNA groups were obviously decreased also(siRNA-1,siRNA-2,siRNA-3 and mixed siRNA groups vs normal or negative control group:0.482±0.035,0.402±0.003.0.479±0.026 and 0.446±0.017 vs 0.826±0.031 or 0.835±0.024,P＜0.01).Conclusion CDC20 siRNA has remarkable inhibitory effect on the expression of CDC20 mRNA and protein in mouse Hepal-6 cell lines.
