微小RNA分子miR-214对食管鳞癌细胞Eca109侵袭能力的影响

Effect of miR-214 on invasive capacity of esophageal squamous carcinoma cell line Eca109

目的探讨微小RNA分子miR-214对食管鳞癌细胞Eca109侵袭能力的影响及其可能的分子机制。方法根据人源miR-214序列合成双链模拟物。通过脂质体转染将miR-214模拟物分子导入食管鳞癌Eca109细胞中,转染无关miRNA模拟物作为对照。采用定量PCR法检测细胞中成熟miR-214分子水平,以Matrigel包被的Transwell实验测定侵袭细胞率,通过蛋白质印迹法检测细胞中E-钙黏蛋白(E-cadherin)的表达,利用流式细胞术检测E-cadherin阳性细胞率。结果 Eca109细胞转染miR-214模拟物48h后,其成熟miR-214水平较对照组明显上升(P<0.01);细胞侵袭能力较对照组降低(P<0.05)。同时,miR-214转染组的E-cadherin表达水平及E-cadherin阳性细胞率较对照组下降(P<0.05)。结论 miR-214可能通过抑制细胞上皮间质转化(EMT)的方式抑制食管鳞癌细胞的侵袭能力。

Objective To investigate the effect of miR-214 on the invasive capacity of esophageal squamous carcinoma cell line Ecal09, and to explore the possible molecular mechanism. Methods We prepared miR-214 double-stranded mimic and transfected it into Ecal09 cells with Lipofectamine 2000. Ecal09 cells transfected with nonsense miRNA mimics were taken as control. The expression 0f mature miR-214 was determined by qPCR. The capacity of cell invasion was determined by Matrigel-coated Transwell assay. E-cadherin protein expression and the percentage of E-cadherin positive cells were examined by Western blotting analysis and flow cytometry, respectively. Results The expression level of mature miR-214 in the miR-214 mimic transfection group was significantly higher than that in the control group 48 h after transfection （P〈0. 01）. Ecal09 cells transfected with miR-214 mimic showed a significantly lower cell invasive capacity compared to that of cells transfected with control miRNA mimic （P〈O, 05）. Moreover, Ecadherin protein expression and the ratio of E-cadherin positive cells in miR-214 mimic transfection group were both significantly lower than those in the control group （P〈0.05）. Conclusion Our data suggest that miR-214 may inhibit the invasive capacity of esophageal squamous carcinoma cells by repressing the epithelial-mesenchymal transition.