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目的:研究婆罗双树样基因2(sal-like gene 2,SALL2)干扰对宫颈癌He La细胞增殖和凋亡的影响。方法:设计针对SALL2基因的3条小干扰RNA(si RNA-1、2、3),并设载体组及未转染对照组,使用脂质体转染He La细胞。采用逆转录RT-PCR和Western blot检测转染后He La细胞中SALL2 m RNA和蛋白的表达以筛选沉默效果最好的si RNA片段。将最适si RNA片段转染He La细胞后,采用CCK-8检测细胞增殖率;流式细胞术检测细胞周期和凋亡率。结果:对照组He La细胞高表达SALL2。载体组和转染si RNA-3组SALL2的m RNA和蛋白表达与对照组相比,差异均无统计学意义(P均>0.05),而si RNA-1和si RNA-2转染48 h后,He La细胞SALL2的m RNA和蛋白表达均降低,差异均有统计学意义(P均<0.05),其中si RNA-1的沉默效率更好,故后续选择si RNA-1作为干扰组。与对照组相比,转染si RNA-1 48和72 h后细胞的增殖率均升高,差异均有统计学意义(P<0.05);转染48 h后si RNA-1组处于G0/G1期的细胞比例减少,凋亡率明显降低(P均<0.05)。结论:SALL2基因干扰可明显提高He La细胞的增殖率,并降低其凋亡率。提示SALL2基因对宫颈癌He La细胞的增殖有抑制作用。
Objective: To investigate the effect of sal-like gene 2 (SALL2) on the proliferation and apoptosis of cervical cancer HeLa cells. Methods: Three small interfering RNAs (si RNA-1,2,3) targeting SALL2 gene were designed and transfected into HeLa cells by lipofectamine. The expression of SALL2 mRNA and protein in HeLa cells was detected by reverse transcription RT-PCR and Western blot to screen the si RNA fragment with the best silencing effect. After transfection of the optimal siRNA fragment into HeLa cells, the cell proliferation rate was detected by CCK-8. The cell cycle and apoptosis rate were detected by flow cytometry. Results: The control group He La cells highly expressed SALL2. The mRNA and protein expressions of SALL2 in vector group and si RNA-3 transfected group were not significantly different from those in control group (all P> 0.05), while the si RNA-1 and si RNA-2 transfected S48 cells for 48 h , The mRNA and protein expression of SALL2 in HeLa cells both decreased (all P <0.05). Among them, si RNA-1 had better silencing efficiency, so si RNA-1 was selected as the interference group later. Compared with the control group, the proliferation rates of si RNA-1 48 and 72 h after transfection were significantly increased (P <0.05). At 48 h after transfection, si RNA-1 group was at G0 / The proportion of cells in G1 phase decreased and the apoptosis rate decreased significantly (all P <0.05). Conclusion: The SALL2 gene interference can significantly increase the He La cell proliferation rate, and reduce the apoptosis rate. Suggesting that SALL2 gene has an inhibitory effect on the proliferation of cervical cancer HeLa cells.