上调NDRG1表达降低奥沙利铂耐药结肠癌细胞的存活率

Up-regulating NDRG1 expression decrease the survival rate of oxaliplatin-resistant colon cancer cells

目的观察上调NDRG1表达对结肠癌细胞及奥沙利铂耐药结肠癌细胞的毒性作用并探讨其机制。方法以重组真核表达质粒pEGFP-NDRG1-N3转染HCT 116及奥沙利铂耐药的OHP-HCT 116结肠癌细胞,以实时定量(qRT)-PCR法和Western blot法检测转染效率。MTT法检测奥沙利铂对不同结肠癌细胞的毒性及验证OHP-HCT 116细胞的耐药性。给予奥沙利铂干预转染pEGFP-NDRG1-N3的HCT 116细胞及OHP-HCT 116细胞,MTT法检测细胞存活率,流式细胞术检测细胞凋亡率,Western blot法检测凋亡蛋白Bcl-2及p53蛋白水平变化。结果不同浓度奥沙利铂干预下的3株结肠癌细胞中HCT 116细胞对奥沙利铂最敏感,OHP-HCT 116细胞对奥沙利铂耐药得到验证。以不同浓度梯度奥沙利铂干扰转染pEGFP-NDRG1-N3的HCT 116细胞及OHP-HCT 116细胞,与MOCK及siNC组比较,在HCT 116细胞及OHP-HCT 116细胞,转染pEGFP-NDRG1-N3细胞的存活率下降(P<0.05),凋亡率升高(P<0.05),Bcl-2表达降低(P<0.05),p53表达升高(P<0.05)。结论上调NDRG1表达通过调控p53表达改善结肠癌细胞奥沙利铂耐药,提高化疗敏感性。

Objective To observe the toxicity of up-regulating NDRG1 to colon cancer cells and oxaliplatin-resistant colon cancer cells and discuss the mechanism.Methods The recombination eukaryote expression plasmid pEGFP-NDRG1-N3 were transfected into colon cancer cells HCT 116 and oxaliplatin resistant OHP-HCT 116 cell;quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were used to detected the transfection efficiency.The toxicity of oxaliplatin to different colon cancer cells and OHP-HCT 116 oxaliplatin resistance were detected by MTT assay.HCT 116 and OHP-HCT 116 cells were transfected with pEGFP-NDRG1-N3 and interfered with oxaliplatin;cell survival rate and apoptosis were detected by MTT assay and flow cytometry assay respectively;Western blot was used to detect the Bcl-2 and p53 protein expression.Results HCT 116 was most sensitive in three colon cancer cell lines which were interfered with oxaliplatin in different concentrations and OHP-HCT116 cells resistant to oxaliplatin was identified.HCT 116 and OHP-HCT 116 cells which were transfected with pEGFP-NDRG1-N3 were interfered with oxaliplatin in different concentration,and compared with MOCK and siNC groups,the survival rates were decreased in HCT 116 and OHP-HCT 116 cells transfected with pEGFP-NDRG1-N3(P<0.05),while the apoptosis rates were increased(P<0.05);Bcl-2 expression was decreased(P<0.05),while p53 expression was increased(P<0.05).Conclusion Up-regulating NDRG1 expression improves the oxaliplatin resistance to colon cancer cell via regulating p53 expression and enhances chemosensitivity.