miR-143通过负向调控Bcl-2抑制食管癌细胞的增殖

目的探讨miR-143通过负向调控B淋巴细胞瘤-2(Bcl-2)抑制食管癌细胞的增殖。方法 RTPCR法检测食管癌细胞TE-1、EC109、EC9706、KYSE150、KYSE510、SEG-1及人正常食管上皮细胞HEEC中miR-143表达,使用脂质体Lipofectamine~(TM) 2000将miR-143 mimics和miR-143 NC转入EC9706细胞中,48 h后,RT-PCR法检测miR-143表达,CCK-8法检测细胞活力,Ed U染色检测细胞增殖情况,流式细胞术检测细胞周期,RT-PCR及Western blot检测Bcl-2蛋白及mRNA的表达。结果 miR-143在食管癌细胞TE-1,EC109,EC9706,KYSE150,KYSE510及SEG-1中的表达量[(1.36±0.13),(1.08±0.10),(0.89±0.09),(0.95±0.09),(1.32±0.14),(0.96±0.11)]显著低于miR-143在人正常食管上皮细胞HEEC(2.38±0.15)中的表达量(P0.01)。与miR-143 NC组比较,miR-143 mimics组miR-143表达量显著升高(P0.01),细胞活力下降(P0.01),细胞增殖能力降低(P0.01),细胞周期阻滞在G1期(P0.01),同时Bcl-2蛋白及mRNA表达量显著降低(P0.01)。结论miR-143过表达能通过下调Bcl-2表达抑制EC9706细胞增殖。

Inhibition of proliferation of esophageal cancer cells by miR-143 through negative regulation of Bcl-2 expression

Objective To investigate the effect of miR-143 on proliferation of esophageal cancer cells by negative-regulation of B-cell lymphoma-2 (Bcl-2). Methods The expression levels of miR-143 in the esophageal cancer cells TE-1, EC109, EC9706, KYSE150, KYSE510 and SEG-1 and human normal esophageal epithelial cells HEEC were detected by RT-PCR. miR-143 mimics and miR-143 NC were transfected into EC9706 cells by Lipofectamine^TM 2000. After 48 h, the expression levels of miR-143 were detected by RT-PCR, the viability and proliferation of the cells were measured using CCK-8 and EdU staining, the cell cycle was analyzed by flow cytometry, and the expression of Bcl-2 protein and mRNA was detected by Western blot and RT-PCR. Results The expression levels of miR-143 in esophageal cancer cells TE-1, EC109, EC9706, KYSE150, KYSE510 and SEG-1[(1.36±0.13),(1.08±0.10),(0.89±0.09),(0.95±0.09),(1.32±0.14) and (0.96±0.11)] were significantly lower than that in the human normal esophageal epithelial cells HEEC (2.38±0.15) (P < 0.01). Compared with the miR-143 NC group, the expression level of miR-143 was significantly increased (P < 0.01), the cell viability and proliferation were decreased (P < 0.01), the cell cycle was arrested in the G1 phase (P < 0.01), and the expression of Bcl-2 protein and mRNA was significantly decreased (P < 0.01) in the miR-143 mimics group. Conclusions Over-expression of miR-143 can inhibit the proliferation of esophageal cancer EC9706 cells through down-regulation of Bcl-2 expression.