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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 investigate the effect of down-regulating the expression of FoxM1 gene in human renal cell carcinoma 786-O cells on the biological behavior of human renal cell carcinoma. Methods The shFoxM1 plasmid was constructed by using FoxM1 interference sequence and short hairpin RNA (shRNA). 786-O cells were divided into shFoxM1 group and control group, respectively transfected shFoxM1, scramble plasmid 48 h. The mRNA and protein expression of FoxM1 were detected by Real-time PCR and Western blot respectively. The formation of clones was observed by plate clone formation assay. The cell cycle and apoptosis rate were detected by flow cytometry. Transwell assay was used to observe the cell migration and invasion Ability, MTT assay cell culture 0,24,48,72 h when the cell absorbance value. Results Compared with the control group, the mRNA and protein expression of FoxM1 in shFoxM1 group decreased, the rate of cell colony formation and cell absorbance decreased, the proportion of cells in G1 phase increased, the number of G2 and S phase decreased, the early apoptotic rate increased, and the migration and invasion of transmembrane Cells decreased, P <0.05. Conclusion The down-regulation of FoxM1 gene expression can inhibit the proliferation, migration and invasion of 786-O cells in renal cell carcinoma, and arrest the cells in G1 phase and promote the early apoptosis of cells.