下调FoxM1基因表达对肾癌786-O细胞生物学行为的影响

Effect of down-regulating FoxM1 expression on biological behavior of renal cancer 786-O cells

目的观察下调肾癌786-O细胞中叉头框转录因子M1(FoxM1)基因的表达对肾癌细胞生物学行为的影响。方法采用FoxM1的干扰序列和短发夹RNA(shRNA)构建shFoxM1质粒。将786-O细胞分为shFoxM1组与对照组,分别转染shFoxM1、scramble质粒48 h。分别用Real-time PCR、Western blot法检测细胞FoxM1 mRNA及蛋白,平板克隆形成实验观察1周细胞克隆形成率,采用流式细胞仪检测细胞周期及细胞凋亡率,Transwell实验观察细胞迁移和侵袭能力,MTT法检测细胞培养0、24、48、72 h时的细胞吸光度值。结果与对照组比较,shFoxM1组FoxM1 mRNA和蛋白表达减少,细胞克隆形成率、细胞吸光度值降底,G1期细胞比例增加而G2、S期减少,早期凋亡率增加,迁移、侵袭的穿膜细胞减少,P均<0.05。结论下调FoxM1基因表达,可抑制肾癌786-O细胞的增殖、迁移、侵袭能力,使细胞阻滞于G1期,促进细胞的早期凋亡。

Objective To observe the down-regulation of forkhead box protein M 1 （ FoxM1 ） gene expression and to investigate its effect on the biological behavior of human renal cancer cell line 786-O.Methods The shFoxM1 plasmid was constructed by RNA interference sequence and shRNA .786-O cells were divided into the shFoxM 1 group and the con-trol group which were transfected with shFoxM 1 plasmid and scramble plasmid for 48 h, respectively .The expression of FoxM1 mRNA and protein was tested by real-time PCR and Western blotting .Then we analyzed the efficiency of plate colo-ny formation in one week by using colony formation assay , and we used flow cytometry to detect the cell cycle and apoptosis of 786-O cells.The migration and invasion abilities were analyzed by Transwell assay , and the OD value in 0, 24, 48 and 72 hours was tested using MTT assay to analyze the activity of proliferation .Results Compared with the control group , the FoxM1 mRNA and protein expression was decreased , the efficiency of plate colony formation and the cell proliferation were decreased, the cells were blocked in G1 phase, cells of G2 phase and S phase reduced, the early apoptosis rate rose and the migration and invasion cells were decreased in the shFoxM 1 group （all 〈0.05）.Conclusions Down-regulating FoxM1 gene expression suppressed the proliferation , migration and invasion abilities of 786-O cells, blocking cells in G1 phase and promoting early apoptosis .