【摘要】 目的 观察长链非编码核糖核酸（LncRNA）信号传导蛋白Sma和Mad相关蛋白同源物5(SMAD5)反义核糖核酸1(SMAD5-AS1)干扰对人食管癌细胞株Eca109增殖的影响，并初步探讨可能的机制。方法 取人食管癌细胞株Eca109培养传代，分别转染SMAD5-AS1 siRNA沉默质粒、阴性对照质粒，并记为siRNA-1组、siRNA阴性对照组，另取未转染细胞记为对照组，每组设置3个复孔。实时逆转录聚合酶链反应（RT-qPCR）检测各组SMAD5-AS1、表观遗传诱导LncRNA-1(EPIC1)信使核糖核酸（mRNA）表达；细胞计数试剂-8(CCK-8)法检测各组细胞增殖活性；流式细胞仪检测各组细胞周期分布；RT-qPCR检测各组细胞细胞周期D1(cyclinD1)、促凋亡基因（Bad）、抗凋亡基因（Bcl-2）mRNA表达；蛋白质免疫印迹法（WB）检测各组细胞cyclinD1、Bad、Bcl-2蛋白表达。结果 各组细胞SMAD5-AS1、EPIC1 mRNA表达水平比较：对照组>siRNA阴性对照组>siRNA-1组，差异有统计学意义（P<0.05）；各组细胞增殖活性比较：siRNA-1组>siRNA阴性对照组>对照组，差异有统计学意义（P<0.05）；与对照组和siRNA阴性对照组比较，siRNA-1组cyclinD1和Bcl-2 mRNA与蛋白表达均下降，差异有统计学意义（P<0.05）,G1/S期细胞占比、Bad mRNA及蛋白表达均升高，差异有统计学意义（P<0.05），上述指标siRNA阴性对照组和对照组比较差异均无统计学意义（P>0.05）。结论 SMAD5-AS1干扰可抑制人食管癌细胞株Eca109增殖，将其阻滞于G1/S期，推测与下调EPIC1、cyclinD1及Bcl-2表达，上调Bad表达有关。更多还原

【Abstract】 Objective To observe the effect of long chain noncoding RNA(LncRNA)signal transduction protein Sma and Mad related protein homologue 5(SMAD5)antisense RNA 1(SMAD5-AS1)interference on the proliferation of human esophageal cancer cell line Eca109,and to explore its signal pathway regulation mechanism. Methods Human esophageal cancer cell line Eca109 was cultured and subcultured,and transfected with SMAD5-AS1 siRNA silencing plasmid and negative control plasmid,respectively,and recorded as the siRNA-1 group and the siRNA-negative control group. In addition,untransfected cells were recorded as the control group,and 3 multiple wells were set in each group. The messenger ribonucleic acid(mRNA) expressions of SMAD5-AS1 and epigenetic lncRNA-1(EPIC1) were detected by reverse transcription polymerase chain reaction(RT-qPCR). Cell proliferation activities were detected by cell counting kit-8(CCK-8)assay. Cell cycle distributions of each group were detected by flow cytometry. The mRNA expressions of cyclinD1,pro-apoptotic gene(Bad)and anti-apoptotic gene(Bcl-2)were detected by RT-qPCR. The protein expressions of cyclinD1,Bad and Bcl-2 were detected by western blotting(WB). Results Comparison of SAMD5-AS1 and EPIC1 mRNA expression levels in each group:the control group > the siRNAnegative control group > the siRNA-1 group,the difference was statistically significant(P<0.05). Comparison of cell proliferation activity in each group:the siRNA-1 group > the siRNA-negative control group > the control group,the difference was statistically significant(P<0.05). Compared with the control group and the siRNAnegative control group,the expressions of cyclinD1 and Bcl-2 mRNA and protein in the siRNA-1 group were decreased,and the differences were statistically significant(P<0.05). The proportions of cells in G1/S phase and expressions of Bad mRNA and protein were increased(P<0.05)in the siRNA-1 group,and there was no statistical significance in the above indicators between the siRNA negative control group and the control group(P>0.05). Conclusion SMAD5-AS1 interference can inhibit the proliferation of human esophageal cancer cell lines Eca109 and block them in G1/S phase,which is presumed to be related to down-regulation of EPIC1,cyclinD1 and Bcl-2 expressions and up-regulation of Bad expression.更多还原