MicroRNA-21调控口腔鳞癌细胞顺铂耐药及其作用机制  

Expressions of Micro RNA-21 in Oral Squamous Cell Carcinoma and Its Correlation with Drug Resistance by Targeting PTEN

作　　者：陈建萍 王萍 杨庚萍 欧阳少波[3] 黄自坤[4] 

机构地区：[1]江西省萍乡市湘东区人民医院口腔科,萍乡337055 [2]江西省萍乡市湘东区中医院口腔科,萍乡337016 [3]南昌大学附属口腔医院修复科,江西南昌330006 [4]南昌大学第一附属医院检验科,江西南昌330006

出　　处：《实用中西医结合临床》2015年第12期14-18,共5页Practical Clinical Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的:探讨micro RNA-21(mi RNA-21)在人口腔鳞癌顺铂耐药细胞中的表达及其作用机制。方法 :采用实时荧光定量PCR(RT-PCR)法检测mi RNA-21在口腔鳞癌细胞株Tca8113及顺铂耐药细胞株Tca8113/DDP的表达;采用体外转染法将mi RNA-21模拟物(mimics)或抑制物(inhibitor)分别转染Tca8113和Tca8113/DDP细胞,采用RT-PCR检测转染前后细胞中mi RNA-21的表达情况;采用CCK8实验检测转染前后细胞对顺铂敏感性的变化;并用Western blot检测转染前后细胞中PTEN的表达变化。采用双荧光素酶报告基因验证mi RNA-21是否作用于PTEN基因的3’-UTR区预测靶位。结果:mi RNA-21在Tca8113/DDP细胞中的表达水平是Tca8113细胞的(8.26±1.37)倍(P<0.01)。Tca8113细胞转染mi RNA-21 mimics后,细胞中mi RNA-21表达水平是转染前的(10.51±2.18)倍(P<0.01),顺铂对Tca8113细胞增殖的抑制率较转染前明显下降(P<0.05),细胞内PTEN表达水平较转染前明显下降(P<0.05)。Tca8113/DDP细胞在转染mi RNA-21 inhibitor后,细胞中mi RNA-21表达水平是转染前的(0.32±0.14)倍(P<0.01),顺铂对Tca8113/DDP细胞增殖的抑制率与转染前比较明显增加(P<0.05),细胞内PTEN表达水平较转染前明显增高(P<0.05)。经双荧光素酶报告基因验证PTEN是mi RNA-21的靶基因。结论:mi RNA-21在口腔鳞癌顺铂耐药细胞Tca8113/DDP中异常高表达,下调其表达可部分逆转细胞耐药性,mi RNA-21可能通过作用于PTEN参与口腔鳞癌细胞顺铂耐药的发生和发展。Objective： To investi gate the expression of micro RNA-21（mi RNA-21） in cisplatin-resistant oral squamous cell carcinoma cells and its mechanism. Methods： Real-time PCR（RT-PCR） was used to detect the expression of mi RNA-21 in cisplatin-resistant oral squamous cell carcinoma cell line Tca8113/DDP and its parent cell line Tca8113. mi RNA-21 mimics and mi RNA-21 inhibitor in vitro was transiently transferred into Tca8113 and Tca8113/DDP cells respectively. The expression level of mi RNA-21 was detected by RT-PCR. Cell viability was analyzed by CCK8 assay. Expression of epithelial membrane protein-1（PTEN）protein was detected by Western blot. Bioinformatics software predicted the potential target genes of mi RNA-21 and dual luciferase reporter gene was used to analyze the binding between mi RNA-21 and 3＇-UTR of PTEN. Results： The expression level of mi RNA-21 was up-regulated in Tca8113/DDP cells as compared with Tca8113 cells（P〈0.01）. The relative expression level of mi RNA-21 in mi RNA-21mimics-transfected Tca8113 was significantly increased. Over-expression of mi RNA-21 inhibited cisplatin chemosensitivity（P〈0.05）,and the up-regulation of mi RNA-21 led to a significant decrease in the PTEN protein levels（P〈0.05）. The relative expression 1evel of mi RNA-21 in mi RNA-21 inhibitor-transfected Tca8113/DDP was significantly decreased. Low-expression of mi RNA-21 promoted cisplatin chemosensitivity（P〈0.05）, and the down-regulation of mi RNA-21 led to a significant increase in the PTEN protein levels（P〈0.05）. Dual luciferase reporter gene assay indicated that mi RNA-21 regulated PTEN expression by binding to the 3＇-UTR of PTEN m RNA. Conclusions： The expression of mi RNA-21 was up-regulated in Tca8113/DDP cells. Down-regulation the expression of mi RNA-21 could partially reverse the cisplatin-resistance. mi RNA-21 may play a vital role in oral squamous cell carcinoma drug resistance by targeting PTEN.

关 键 词：口腔鳞癌 微小RNA-21 顺铂 耐药 人第10号染色体缺失的磷酸酶 

分 类 号：R739.8[医药卫生—肿瘤]