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目的研究微小RNA106b(miR-106b)对食管癌KYSE150细胞增殖、迁移、侵袭及上皮间质转化(Epithelial-Mesenchymal Transition,EMT)的影响,为临床治疗食管癌提供新的理论依据。方法用miR-106b慢病毒转染KYSE150细胞,实验分为3组:control组(未转染miR-106b的KYSE150细胞)、miR-NC组(转染随机序列)和miR-106b组(转染miR-106b的KYSE150的细胞)。采用实时荧光定量聚合酶联反应(qRT-PCR)和免疫印迹(Western Blot)检测miR-106b和EMT的相关标记物E-钙黏蛋白(E-Cadherin)、神经型钙黏蛋白(N-Cadherin)的表达情况,四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞的增殖能力,划痕实验检测细胞的迁移能力,Transwell方法检测细胞的侵袭能力。结果慢病毒稳定转染miR-106b后,食管癌细胞的增殖、侵袭和迁移能力均增强(P<0.05);qRT-PCR与Western blot检测显示miR-106b组E-Cadherin表达较miR-NC组、control组明显降低(P<0.05);N-Cadherin表达明显升高(P<0.05)。结论 miR-106b能够促进食管癌细胞的增殖、迁移和侵袭,同时能促进上皮间质的转化。
Objective To investigate the effect of miR-106b on the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of esophageal cancer KYSE150 cells and to provide a new theoretical basis for clinical treatment of esophageal cancer. Methods KYSE150 cells were transfected with miR-106b lentivirus. The experiment was divided into 3 groups: control group (KYSE150 cells without miR-106b), miR-NC group (transfected with random sequence) and miR-106b group miR-106b cells of KYSE150). Real-time quantitative PCR (qRT-PCR) and Western Blot were used to detect the expressions of E-Cadherin, N-Cadherin and miR- ). The MTT method was used to detect the proliferation of cells. Scratch assay was used to detect the migration ability of cells. Transwell method was used to detect the invasion ability of cells. Results The results of qRT-PCR and Western blot showed that the expression of E-cadherin in miR-106b group was significantly higher than that in miR-106 group (P <0.05) , While the control group was significantly lower (P <0.05); the expression of N-Cadherin was significantly higher (P <0.05). Conclusion miR-106b can promote the proliferation, migration and invasion of esophageal cancer cells and promote the transformation of epithelial-mesenchymal cells.