【摘要】 目的 研究长链非编码(Lnc)RNA ANO1-AS1对食管鳞癌细胞迁移和侵袭的影响及其可能的分子机制。方法 食管癌细胞株(EC109、TE)转染ANO1-AS1敲减慢病毒，沉默ANO1-AS1表达为LV-sh-ANO1-AS1组，阴性肿瘤对照为LV-Con组，未进行转染为空白对照(Blank)组。通过实时荧光定量聚合酶链反应(qRT-PCR)检测各组食管癌细胞中ANO1-AS1和ANO1的相对表达量。通过划痕实验和Transwell迁移实验检测各组细胞的迁移能力。通过Transwell侵袭实验检测各组细胞的侵袭能力。通过Western印迹测定每组细胞中ANO1、基质金属蛋白酶(MMP)2、MMP9、上皮间质转化(EMT)相关蛋白及细胞外信号调节激酶(ERK)/丝裂原活化蛋白激酶(MAPK)信号通路相关蛋白表达。结果 qRT-PCR结果显示，转染ANO1-AS1敲减慢病毒后，食管癌细胞EC109、TE中LV-sh-ANO1-AS1组LncRNA ANO1-AS1和ANO1 mRNA表达水平均显著低于LV-Con组和Blank组(P<0.05)。划痕实验结果表明，LV-sh-ANO1-AS1组细胞划痕愈合率显著低于LV-Con组和Blank组(P<0.05)。Transwell结果表明，LV-sh-ANO1-AS1组细胞迁移和侵袭能力显著低于LV-Con组和Blank组(P<0.05)。Western印迹结果显示，与LV-Con组和Blank组比较，LV-sh-ANO1-AS1组MMP2、MMP9、Vimentin、p-ERK及ANO1蛋白表达水平显著降低，E-cadherin蛋白表达水平显著升高(P<0.05)。结论 LncRNA ANO1-AS1在食管鳞癌细胞EC109、TE中高表达，并可能通过影响ERK/MAPK信号通路促进食管癌细胞的迁移和侵袭。更多还原

【Abstract】 Objective To study the effect of long non-coding(Lnc)RNA ANO1-AS1 on migration and invasion of esophageal squamous cell carcinoma cells and its possible molecular mechanism.Methods ANO1-AS1 knockdown virus was transfected into esophageal cancer cell lines(EC109, TE). The ANO1-AS1 expression silencing was named LV-sh-ANO1-AS1 group, the negative tumor control was LV-Con group and untreated cells were blank control(Blank) group. The relative expression levels of ANO1-AS1 and ANO1 in each group of esophageal cancer cells were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR). The scratch test and Transwell migration test were used to detect the migration ability of cells in each group. The Transwell invasion test was used to detect the invasion ability of cells in each group. The expressions of ANO1, matrix metalloproteinases(MMP)2, MMP9, epithelial mesenchymal transitions(EMT) related proteins and extracellular regulated protein kinases(ERK)/mitogen-activated protein kinase(MAPK) signal pathway related proteins in each group of cells were detected by Western blot.Results qRT-PCR results showed that after transfection with ANO1-AS1 knockdown virus, the expression levels in esophageal cancer cell EC109, TE of LncRNA ANO1-AS1 and ANO1 mRNA in LV-sh-ANO1-AS1 group were significantly lower than those in LV-Con group and Blank group(P<0.05). The results of cell scratch test showed that the healing rate of cell scratch in LV-sh-ANO1-AS1 group was significantly slower than that in LV-Con group and Blank group(P<0.05). The results of Transwell experiment showed that the migration and invasion ability of cells in LV-sh-ANO1-AS1 group were significantly reduced compared with LV-Con group and Blank group(P<0.05). Western blot results showed that compared with LV-Con group and Blank group, the expression levels of MMP2, MMP9, Vimentin, p-ERK and ANO1 protein were significantly decreased(P<0.05), and the expression level of E-cadherin protein was significantly increased in LV-sh-ANO1-AS1 group(P<0.05). Conclusions LncRNA ANO1-AS1 is highly expressed in esophageal squamous cell carcinoma cells EC109 and TE, and it might promote the migration and invasion of esophageal cancer cells by regulating ERK/MAPK signal pathway.更多还原