siRNA靶向沉默fbxl20基因对肝癌SMMC-7721细胞生物学行为的影响

目的:通过沉默fbxl20基因,以观察基因沉默后对SMMC-7721细胞的增殖、迁移和周期分布的影响。方法:通过脂质体Lipofectamine2000介导转染SMMC-7721细胞,靶向fbxl20基因的siRNA片段阻低肝癌SMMC-7721细胞株fbxl20 mR-NA表达。结果:siRNA能有效沉默SMMC-7721细胞fbxl20 mRNA的表达,沉默效率为71.49%。MTT试验实验显示:siRNA转染SMMC-7721细胞后,细胞增殖能力显著降低。Transwell小室和划痕愈合实验表明,fbxl20基因被沉默后,SMMC-7721细胞在迁移能力明显减弱,迁移抑制率高达62.03%。流式细胞术检测细胞周期结果显示:转染48 h后,SMMC-7721实验组细胞G1/G0期比率(81.21%)显著高于阴性对照组(71.49%)和空白对照组(67.90%)。结论:本实验运用RNA干扰技术成功抑制了SMMC-7721细胞中fbxl20基因的表达,并且发现fbxl20基因可能与肝癌细胞的增值和迁移相关。

Effect of siRNA targeted to fbxl20 on biological behavior of liver cancer SMMC-7721 cells

Objective: To study the effect of siRNA targeted to F-box and leucine-rich repeat protein 20(fbxl20) on fbxl20 mRNA level of expression,proliferation capacity,migration ability,and the cell cycle of SMMC-7721 cancer cells.Methods:Lipofectamine 2000 was employed as the transfecting reagent to transport the siRNA into the SMMC-7721 cancer cells.Results:It was shown that the fbxl20 level of expression was significantly decreased after RNAi,and the inhibition rate was 71.49%.MTT assay showed that the proliferation activity was significantly reduced after RNAi.It was found that the migration ability of SMMC-7721 was decreased after RNAi through transwell chamber assay and scratch healing experiment.The inhibition rate was 62.03%.The flow cytometry revealed that the G1/G0 rate(81.21%)of SMMC-7721 was significantly increased after RNAi,compared to theat of the negative control(71.49%) and the blank control group(67.90%).Conclusion: It is concluded that both the decreased proliferation capacity,migration ability,and the increased G1/G0 rate of SMMC-7721 are closely related with the silencing of the fbxl20 expression.