过表达miR-218增强食管鳞状细胞癌对顺铂敏感性

目的 研究miR-218 在食管鳞癌细胞中对顺铂药物敏感性的影响.方法 采用递增药物浓度、间歇冲击作用的方法构建耐药细胞株;实时荧光定量PCR检测miR-218 在食管鳞癌细胞株Eca109 与顺铂耐药株Eca109-CisR差异性表达;进一步在顺铂耐药株Eca109-CisR中过表达miR-218:① CCK8实验检测细胞增殖;② 流式细胞仪检测细胞凋亡率;③ Western blot方法检测凋亡抑制蛋白Survivin、Mcl-1及Bcl-2的相对表达量.结果 miR-218在顺铂耐药株Eca109-CisR细胞中呈低表达;过表达 miR-218 增加顺铂耐药株Eca109-CisR对顺铂的药物敏感性;Western blot结果证实凋亡相关蛋白Survivin、Mcl-1 及 Bcl-2相对表达减少.结论 miR-218表达有助于增加食管鳞癌细胞对顺铂的敏感性,且给对顺铂耐药的食管鳞癌患者提供潜在的基因治疗靶点.

Overexpression of miR-218 enhances the chemosensitivity of esophageal squamous cell carcinoma to cisplatin

Objective To investigate the role of miR-218 in cisplatin sensitivity of esophageal cancer cells. Meth-ods Adopt the method of increasing drug concentrations, intermittent impact construction of drug resistant cell lines. Quantitative real-time polymerase chain reaction ( qPCR) was carried out to analyze miR-218 expression in human esophageal cancer cell line Eca109 and a cisplatin-resistant subline ( ECa109-CisR cells) . The effects of miR-218 transfection on ECa109-CisR cell viability, including cell viability and apoptosis rate were confirmed using CCK8 assay, or flow cytometry, respectively. Western blot was used to determine the relative expression of apopto-sis inhibitoryprotein Survivin,Mcl-1 and Bcl-2. Results We found that miR-218 was significantly decreased in ECa109-CisR cells compared with parent Eca109 cells. Overexpression of miR-218 by mimics’ transfection would enhance cisplatin sensitivity evaluated by cell viability inhibition and apoptosis promotion. Western blot results con-firmed the decreased expression of apoptosis-related proteins Survivin, Mcl-1, and Bcl-2. Conclusion miR-218 expression contributes to the increased sensitivity of esophageal squamous carcinoma cells to cisplatin and provides a potential gene therapy target for cisplatin-resistant esophageal squamous cell carcinoma patients.