miRNA-181a在人肺腺癌细胞中的表达及对细胞功能的影响  

Expression of mi RNA-181a in human lung adenocarcinoma cells and its effect on cell function

作　　者：赵铎[1] 金柱[1] 宋亚亚[1] 高宝安[1] 

机构地区：[1]三峡大学呼吸病研究所,宜昌市中心人民医院呼吸科,湖北宜昌443003

出　　处：《中国病理生理杂志》2016年第8期1395-1402,共8页Chinese Journal of Pathophysiology

摘　　要：目的:研究微小RNA-181a(mi RNA-181a)在不同人肺腺癌细胞中的表达及其转染人肺腺癌耐药细胞A549/DDP后对其细胞功能的影响及机制。方法:利用实时荧光定量PCR方法检测mi RNA-181a在人正常肺上皮细胞系BEAS-2B、人肺腺癌细胞系A549、人肺腺癌耐药细胞系A549/DDP中的表达;利用p Genesil-mi RNA-181a真核表达质粒转染A549/DDP细胞,同时设置未转染组和空载组;分别采用实时荧光定量PCR、MTT法、流式细胞术、Transwell实验以及Western blot法检测mi RNA-181a转染前后的表达情况、对A549/DDP细胞活力、细胞周期、细胞侵袭能力和顺铂(DDP)作用下的细胞生长抑制率、细胞凋亡率的影响以及对A549/DDP细胞中mi RNA-181a的靶基因bcl-2和p53蛋白表达的影响。结果:mi RNA-181a在A549和A549/DDP中的表达量显著低于BEAS-2B(P<0.05),且在A549/DDP中表达量最低;mi RNA-181a转染A549/DDP细胞后其表达显著升高(P<0.05)且能够抑制A549/DDP细胞活力、细胞周期和细胞侵袭能力(P<0.05),同时升高DDP作用下A549/DDP细胞的生长抑制率和细胞凋亡率(P<0.05);mi RNA-181a转染A549/DDP细胞后抑制Bcl-2蛋白的表达而促进P53蛋白的表达(P<0.05)。结论:mi RNA-181a可能参与了肺腺癌的发生发展,mi RNA-181a可作为人肺腺癌治疗的新靶点。AIM： To study the expression of micro RNA（mi RNA）-181 a in different human lung adenocarcinoma cell lines,and to investigate the effect of mi RNA-181 a on cell function and its mechanism in human lung adenocarcinoma drug resistant cell A549 / DDP. METHODS： Real-time PCR was used to detect the expression of mi RNA-181 a in BEAS-2B cells,A549 cells and A549 / DDP cells. The A549 / DDP cells were transfected with p Genesil-mi RNA-181 a eukaryotic expression plasmid. At the same time,the untransfection group and negative transfection group were also set up. The expression of mi RNA-181 a,cell viability,cell growth inhibition and apoptosis rate during cis-diamminedichloroplatinum（ DDP）treatment,cell cycle,cell invasion,the protein expression of mi RNA-181 a target genes bcl-2 and p53 in the A549 / DDP cells were determined by real-time fluorescence quantitative PCR,MTT assay,flow cytometry,Transwell method and Western blot,respectivly. RESULTS： The expression of mi RNA-181 a in A549 cells and A549 / DDP cells was significantly lower than that in BEAS-2B cells,and the lowest expression level was observed in A549 / DDP cells（ P 0. 05）. The expression of mi RNA-181 a in A549 / DDP cells was significantly increased after transfection with p Genesil-mi RNA-181a（ P 0. 05）. The cell viability,cell cycle and invasion ability of the A549 / DDP cells were inhibited after mi RNA-181 a transfection（ P 0. 05）. The cell growth inhibition rate and apoptotic rate of the A549 / DDP cells were increased（ P 0. 05）. The expression of Bcl-2 was reduced,but the expression of P53 was increased after transfection with mi RNA-181 a in A549 / DDP cells（ P 0. 05）. CONCLUSION： mi RNA-181 a may be correlated with the development of human lung adenocarcinoma. mi RNA-181 a can serve as a new target for treatment of lung cancer.

关 键 词：微小RNA-181a A549/DDP细胞系 细胞周期 细胞凋亡 细胞侵袭 

分 类 号：R730.23[医药卫生—肿瘤]