摘要
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 CDC20mRNA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48h后，采用逆转录-聚合酶链反应（RT-PCR）和WesternNot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、25nmol／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±0．031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。
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 Hepa1-6 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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第1期47-49,共3页
Chinese Journal of Experimental Surgery
基金
基金项目：深圳市科技计划医疗卫生重点项目（200507）
