沉默Ambra1基因增强大肠癌细胞对5-FU的化疗敏感性

siRNA-Ambra1 Enhances Chemosensitivity of Human Colon Cancer Cells for 5-FU

目的:观察5-FU对沉默Ambra1基因的大肠癌细胞的化疗敏感性影响。方法:将靶向Ambra1基因的si RNA转染至大肠癌细胞HCT15和HCT116,用Real-time PCR和Western blotting方法检测Ambra1 mRNA和蛋白的表达;采用MTS方法检测HCT15和HCT116细胞的活力。结果:HCT15、HCT116细胞转染siRNA-Ambra1-1、siRNA-Ambra1-2后的Ambra1 mRNA表达水平均分别低于siRNAAmbra1-HCT15-NC、siRNA-Ambra1-HCT116-NC细胞(P<0.05);HCT15、HCT116细胞转染后的蛋白表达均低于阴性对照;4μg/mL时,HCT15和HCT116细胞转染si RNA-Ambra1-1、siRNA-Ambra1-2后的细胞克隆形成数均少于0μg/m L(P<0.05),而且HCT15、HCT116细胞转染si RNA-Ambra1-1、siRNAAmbra1-2后的细胞克隆形成数均分别少于si RNA-Ambra1-HCT15-NC、si RNA-Ambra1-HCT116-NC(P<0.05);HCT15、HCT116细胞转染si RNA-Ambra1-1、siRNA-Ambra1-2后的细胞生存率均分别低于siRNA-Ambra1-HCT15-NC、siRNA-Ambra1-HCT116-NC(P<0.05);且随着药物浓度增加,已转染了siRNA-Ambra1的细胞生存率均低于阴性对照组。结论:沉默Ambral基因可提高大肠癌细胞对5-FU的化疗敏感性。

Objective：To observe the effect of 5-FU on chemosensitivity of colorectal cancer cells silencing Ambra1 gene.Method：si RNA targeting Ambra1 gene was transfected into colorectal cancer cells HCT15 and HCT116.The expressions of Ambra1 m RNA and protein were detected by Real-time PCR and Western blotting.The viability of HCT15 and HCT116 cells was detected by MTS.Result：The expression levels of Ambra1 m RNA in HCT15 and HCT116 cells transfected with si RNA-Ambra1-1 and si RNA-Ambra1-2 were lower than those of si RNA-Ambra1-HCT15-NC and si RNA-Ambra1-HCT116-NC cells respectively（P〈0.05）.The protein expression of HCT15 and HCT116 cells after transfection were lower than those of negative control.When 4 μg/m L, the cell clone formation number of HCT15 and HCT116 cells transfected with si RNA-Ambra1-1 and si RNAAmbra1-2 were less than 0 μg/m L（P〈0.05）,and the cell clone formation number of HCT15 and HCT116 cells transfected with si RNA-Ambra1-1 and si RNA-Ambra1-2 were less than those of si RNA-Ambra1-HCT15-NC and si RNA-Ambra1-HCT116-NC cells respectively（P〈0.05）.The cell survival rates of HCT15 and HCT116 cells transfected with si RNA-Ambra1-1 and si RNA-Ambra1-2 were lower than those of i RNA-Ambra1-HCT15-NC and si RNA-Ambra1-HCT116-NC cells respectively（P〈0.05）,with the increase of drug concentration,the survival rate of the transfected si RNA-Ambra1 cells was lower than that of the negative control group. Conclusion：si RNA-Ambralcan enhance chemotherapy sensitivity of colon cancer cells to 5-FU.