miR-199a-3p在食管鳞癌细胞中的表达及其对细胞增殖和凋亡的影响

Expression of miR-199a-3p and its effects on cell proliferation and apoptosis in esophageal squamous cell carcinoma cells

目的:探讨miR-199a-3p在食管鳞癌细胞中的表达情况及其对细胞增殖、凋亡的影响。方法:采用Realtime PCR法检测miR-199a-3p在5株食管鳞癌细胞TE1、Eca109、EC9706、KYSE450、KYSE790中的表达。用siRNAMateTM转染试剂将人工合成的miR-199a-3p模拟物mimics转染Eca109、EC9706细胞后,采用Real-time PCR法检测miR-199a-3p的表达,采用克隆形成实验、CCK-8法及双染法检测细胞克隆形成能力、增殖能力和凋亡率,Western blot检测m TOR、p-p70S6K和Bcl-2蛋白的表达。以未转染细胞为空白对照,以转染mimics阴性对照的细胞为阴性对照。结果:miR-199a-3p在5株食管鳞癌细胞中均有表达,表达水平差异无统计学意义(P>0.05)。与空白对照组和阴性对照组细胞相比,转染mimics的Eca109和EC9706细胞中miR-199a-3p相对表达量显著升高(P<0.001),细胞克隆形成能力降低(P<0.001),细胞增殖能力降低(P<0.001),细胞凋亡率增加(P<0.001),m TOR、p-p70S6K及Bcl-2蛋白表达水平降低。结论:miR-199a-3p可能是通过调节m TORC1/p70S6K信号通路来抑制食管鳞癌细胞的增殖并促进细胞的凋亡。

Aim： To investigate the expression of miR-199a-3p in esophageal squamous cell carcinoma（ ESCC） cells and its effects on cell proliferation and apoptosis. Methods： The expression of miR-199a-3p in five ESCC cell lines（ TE1,Eca109,EC9706,KYSE450,KYSE790） was detected by Real-time PCR. miR-199a-3p mimics was transfected into Eca109 and EC9706 cells,respectively by siRNA-MateTM,and then the colony capacity,proliferation capacity and apoptosis rate were detected by colony formation assay,CCK-8 assay and Annexin V-FITC and PI staining method,respectively,the expressions of m TOR,p-p70S6 K and Bcl-2 protein were detected by Western blot. Cells without transfection were the blank control,and cells transfected with negative mimics were the negative control. Results： miR-199a-3p expressed in five ESCC cell line cells,and the expression level had no significant difference（ P〈0. 05）. Compared with negative control and blank control,the expression level of miR-199a-3p in Eca109 and EC9706 cells transfected with miR-199a-3p mimics increased（ P〈0. 001）,the colony formation capacity and proliferation capacity decreased（ both P〈0. 001）,the apoptosis rate increased（ P〈0. 001）,and the protein expressions of m TOR,p-p70S6 K and Bcl-2 decreased. Conclusion： miR-199a-3p might inhibit cell proliferation and promote cell apoptosis in ESCC cells through m TORC1/p70S6 K signal pathway.