miR-143通过负向调控Bcl-2抑制食管癌细胞的增殖  

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

作　　者：胡琛琛 张莉[1] 

机构地区：[1]湖北省肿瘤医院重症医学科,武汉430079

出　　处：《中国比较医学杂志》2017年第9期65-70,共6页Chinese Journal of Comparative Medicine

摘　　要：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(P<0.01)。与miR-143 NC组比较,miR-143 mimics组miR-143表达量显著升高(P<0.01),细胞活力下降(P<0.01),细胞增殖能力降低(P<0.01),细胞周期阻滞在G1期(P<0.01),同时Bcl-2蛋白及mRNA表达量显著降低(P<0.01)。结论miR-143过表达能通过下调Bcl-2表达抑制EC9706细胞增殖。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 detec-ted 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 ＆#177; 0. 13）, （1. 08 ＆#177; 0. 10）, （0. 89 ＆#177; 0. 09）, （0. 95 ＆#177; 0. 09）, （1. 32 ＆#177; 0. 14） and （0. 96 ＆#177; 0. 11）] were significantly lower than that in the human normal esophageal epithelial cells HEEC （2. 38 ＆#177; 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 ＆lt;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.

关 键 词：MIR-143 食管癌 EC9706 增殖 B淋巴细胞瘤-2 Bcl-2 

分 类 号：R33[医药卫生—人体生理学]