| 陈建萍 王萍 杨庚萍 欧阳少波 黄自坤.MicroRNA-21调控口腔鳞癌细胞顺铂耐药及其作用机制[J].实用中西医结合临床,2015,15(12):14-18 |
| MicroRNA-21调控口腔鳞癌细胞顺铂耐药及其作用机制 |
| Expressions of MicroRNA-21 in Oral Squamous Cell Carcinoma and Its Correlation with Drug Resistance by Targeting PTEN |
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| DOI： |
| 中文关键词: 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 |
| 英文关键词: Oral squamous cell carcinoma MicroRNA-21 Cisplatin Drug resistance PTEN |
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| 中文摘要: |
| 目的：探讨microRNA-21（miRNA-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miRNA-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miRNA-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miRNA-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miRNA-21是否作用于PTEN基因的3’-UTR区预测靶位。结果：miRNA-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miRNA-21 mimics后，细胞中miRNA-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miRNA-21 inhibitor后，细胞中miRNA-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miRNA-21的靶基因。结论：miRNA-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，miRNA-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。 |
| 英文摘要: |
| Objective: To investigate the expression of microRNA-21 (miRNA-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 miRNA-21 in cisplatin-resistant oral squamous cell carcinoma cell line Tca8113/DDP and its parent cell line Tca8113. miRNA-21 mimics and miRNA-21 inhibitor in vitro was transiently transferred into Tca8113 and Tca8113/DDP cells respectively. The expression level of miRNA-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 miRNA-21 and dual luciferase reporter gene was used to analyze the binding between miRNA-21 and 3’-UTR of PTEN. Results: The expression level of miRNA-21 was up-regulated in Tca8113/DDP cells as compared with Tca8113 cells (P＜0.01). The relative expression level of miRNA-21 in miRNA-21 mimics-transfected Tca8113 was significantly increased. Over-expression of miRNA-21 inhibited cisplatin chemosensitivity (P＜0.05), and the up-regulation of miRNA-21 led to a significant decrease in the PTEN protein levels (P＜0.05). The relative expression 1evel of miRNA-21 in miRNA-21 inhibitor-transfected Tca8113/DDP was significantly decreased. Low-expression of miRNA-21 promoted cisplatin chemosensitivity (P＜0.05), and the down-regulation of miRNA-21 led to a significant increase in the PTEN protein levels (P＜0.05). Dual luciferase reporter gene assay indicated that miRNA-21 regulated PTEN expression by binding to the 3’-UTR of PTEN mRNA. Conclusions: The expression of miRNA-21 was up-regulated in Tca8113/DDP cells. Down-regulation the expression of miRNA-21 could partially reverse the cisplatin-resistance. miRNA-21 may play a vital role in oral squamous cell carcinoma drug resistance by targeting PTEN. |
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