miRNA-25对人食管鳞状细胞癌细胞株TE1增殖的影响  

Effects of miRNA-25 on proliferation of human esophageal squamous-cell carcinoma cell line TE1

作　　者：张伟[1,2] 周勇慧[3] 庞一强[4] 

机构地区：[1]内蒙古科技大学包头医学院病理教研室,内蒙古包头014010 [2]内蒙古科技大学包头医学院第一附属医院病理科,内蒙古包头014010 [3]内蒙古科技大学包头医学院第一附属医院普外科二病区,内蒙古包头014010 [4]包头市第四医院神经外科,内蒙古包头014030

出　　处：《中国病理生理杂志》2015年第11期1979-1985,共7页Chinese Journal of Pathophysiology

摘　　要：目的:探讨过表达/沉默微小RNA-25(microRNA-25,miRNA-25)对人食管鳞状细胞癌细胞株TE1增殖能力的影响及其作用机制。方法:RT-PCR检测各临床样品组织中miRNA-25的表达水平。建立miRNA-25过表达/沉默的TE1细胞株,采用CCK-8法、Brd U实验和流式细胞术检测TE1细胞增殖能力的变化及细胞周期状态;Western blot法和RT-PCR法检测细胞周期调控因子细胞周期蛋白E1(cyclin E1)和细胞周期蛋白依赖性激酶2(CDK2)的蛋白与mRNA表达水平。结果:miRNA-25在食管黏膜组织中具有特异性表达并在TE1细胞中呈高表达。CCK-8法和Brd U实验结果显示过表达miRNA-25的TE1细胞的增殖能力显著增加(P<0.05),而沉默miRNA-25抑制TE1细胞的增殖;流式细胞术检测结果显示过表达miRNA-25可明显促进TE1细胞从G0/G1期向S期转换,沉默miRNA-25则抑制其转换。同时Western blot和RT-PCR实验结果显示过表达miRNA-25后,cyclin E1和CDK2的蛋白和mRNA表达水平均显著增加(P<0.05),沉默miRNA-25后则显著减少(P<0.05)。结论:miRNA-25能够促进人食管鳞状细胞癌细胞株TE1的增殖,其作用机制可能与促进细胞周期转换及上调cyclin E1和CDK2的表达水平有关,提示miRNA-25可作为治疗食管鳞状细胞癌的一个潜在靶点。AIM ： To investigate the effects and mechanisms of microRNA-25 （miRNA-25） on the proliferation of human esophageal squamous-cell carcinoma cell line TEl. METHODS： The abundance of miRNA-25 in different tis- sues was measured by RT-PCR. After silencing or over-expression of miRNA-25 with mimics or inhibitor in TEl cells, the cell proliferation, cell cycle distribution and the expression of cyclin E1 and cyclin-dependent kinase 2 （CDK2） at mRNA and protein levels were measured by CCK-8 assay, BrdU detection, flow cytometry, RT-PCR and Western blot, respective- ly. RESULTS ： miRNA-25 was prominent in esophageal mucosal tissue and highly expressed in TEl cells （P 〈 0. 05 ）. O- ver-expression of miRNA-25 increased TEl cell proliferation, promoted the cell cycle progression and enhanced the en- trance of the cells into S phase （P 〈 0. 05 ）. Inverse results were obtained after down-regulation of miRNA-25 （P 〈 0. 05）. Furthermore, the expression of cyclin E1 and CDK2 at mRNA and protein levels was significantly increased after over-ex- pression of miRNA-25, but decreased after down-regulation of miRNA-25 （P 〈 0. 05 ）. CONCLUSION ： miRNA-25 en- hances cell cycle transition by increasing the expression of cyclin E1 and CDK2, thus accelerating TEl cell proliferation. This study provides a novel mechanism by which miRNA-25 increases the proliferation of human esophageal squamous-cell carcinoma cell line TEl, suggesting that down-regulation of miRNA-25 may be a potential new therapeutic strategy for trea- ting esophageal squamous-cell carcinoma.

关 键 词：微小RNA-25 人食管鳞状细胞癌细胞株TE1 细胞增殖 细胞周期蛋白E1 细胞周期蛋白依赖性激酶2 

分 类 号：R363.14[医药卫生—病理学]