【摘要】 目的:研究靶向RAD18基因的小干扰RNA(siRNA)对食管鳞癌细胞ECA-109增殖和化疗敏感性的影响。方法:合成针对RAD18基因的siRNA(RAD18-siRNA),通过脂质体将其转染人食管鳞癌ECA-109细胞株,采用荧光定量PCR方法检测ECA-109 RAD18和CyclinD1 mRNA的表达,蛋白质印迹法检测其RAD18和CyclinD1蛋白的表达;应用CCK-8法检测ECA-109细胞的增殖和化疗药物对ECA-109细胞存活率的影响,流式细胞术检测ECA-109细胞周期;采用Pearson检验分析RAD18与CyclinD1基因表达的相关性。结果:与未转染组比较,RAD18-siRNA组RAD18表达明显降低(P<0.05)。RAD18-siRNA组细胞增殖抑制(P<0.05),G1期细胞数增多,G2/M期细胞数减少(P<0.05)。不同浓度顺铂或5-氟尿嘧啶处理细胞后,两组细胞的存活率均下降(均P<0.05),但RAD18-siRNA组细胞半数抑制浓度较未转染组减少(P<0.05)。在食管鳞癌组织中,RAD18基因mRNA的表达与CyclinD1基因mRNA的表达呈正相关(r=0.478,P<0.01)。结论:下调RAD18表达能够抑制食管鳞癌细胞的增殖,提高其对化疗药物的敏感性;CyclinD1在食管鳞癌中的表达水平可能参与这一过程。更多还原

【Abstract】 Objective:To investigate the effect of RAD18-siRNA on cell proliferation and chemotherapy sensitivity of esophageal squamous cell carcinoma(ESCC) ECA-109 cells.Methods:RAD18-siRNA was transfected into human ECA-109 cells by Lipofectamine 3000.Quantitative PCR and Western blot were performed to detect RAD18 and CyclinDl expression;CCK-8 assay was used to determine cell proliferation and chemotherapy drug sensitivity;flow cytometry was used to determine cell cycle.Correlation between RAD 18 and CyclinDl mRNA expression was analyzed by Pearson’s correlation.Results:Compared with non-transfected cells,the expression of RAD18 in RAD18-siRNA group was significantly decreased(P <0.05).The cell proliferation was inhibited(P < 0.05) and the cell number of Gl phase was increased,G2/M phase cells decreased(P<0.05) in RAD18-siRNA group.After treatment with different concentrations of cisplatin or 5-FU,the survival rate of the two cell groups was reduced(all P <0.05),and the IC50 of RAD18-siRNA group was significantly lower than that of non-transfected group(P < 0.05).The mRNA expression of RAD18 was positively correlated with CyclinD1 expression in ESCC tissues(r = 0.478,P < 0.01).Conclusion:Down-regulated expression of RAD18 can decrease the cell proliferation and increase chemo-sensitivity of ESCC cells,and CyclinD1 may participate in the process.更多还原