沉默miR-21对子宫内膜癌顺铂耐药细胞株Ishikawa/DDP的影响
| 作者: |
1项锦红,
1丁秋霞,
1黄强,
1杨鹰
1 第三军医大学第二附属医院妇产科，重庆 400000 |
| 通讯: |
项锦红
Email: 15273516@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2017.05.009 |
摘要
目的：观察沉默miR-21对子宫内膜癌顺铂耐药细胞株Ishikawa/DDP的影响。方法：以Lipofectamine 2000介导miR-21抑制剂转染Ishikawa/DDP细胞株，同时设置阴性组和耐药组。采用反转录PCR检测miR-21、多药耐药基因MDR1、促凋亡基因Bax和抗凋亡基因Bcl-2的表达。采用蛋白印迹法检测多药耐药蛋白P-gp、促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达。采用噻唑蓝比色法检测细胞对顺铂的敏感性。采用流式细胞术检测细胞凋亡情况。结果：与耐药组和阴性组比较，抑制剂组miR-21，MDR1和Bcl-2 mRNA表达显著下调(P<0.01)，而Bax mRAN表达显著上调(P<0.001)；抑制剂组P-gp和Bcl-2蛋白表达显著低下调(P<0.05)，而Bax蛋白表达显著上调(P<0.001)。与耐药组和阴性组比较，顺铂对抑制剂组的IC50值显著(P<0.001)；顺铂对抑制剂组细胞的诱导凋亡率显著增加(P<0.001)。结论：沉默miR-21可显著提高Ishikawa/DDP细胞株对顺铂的敏感性，并促进细胞凋亡，其具体机制可能与下调MDR1，P-gp和Bcl-2表达，以及上调Bax表达有关。
关键词：
miR-21
子宫内膜癌
顺铂
耐药
Effect of silencing miR-21 on Ishikawa/DDP cisplatin resistant cell in endometrial carcinoma
CorrespondingAuthor: XIANG Jinhong Email: 15273516@qq.com
DOI: 10.3978/j.issn.2095-6959.2017.05.009
Abstract
Objective: To observe the effects of silencing miR-21 on Ishikawa/DDP cisplatin resistant cell in endometrial carcinoma. Methods: Ishikawa/DDP cisplatin resistant cell was transfected with miR-21 inhibitor by Lipofectamine 2000. At the same time, control group and drug resistant group were set up. The expression of miR-21, MDR1, Bax and Bcl-2 was detected by reverse transcription PCR. The protein expression of P-gp, Bax and Bcl-2 was detected by Western blot. The sensitivity of Ishikawa/DDP cells to cisplatin was detected by MTT. The cell apoptosis was detected by flow cytometry. Results: Compared with the drug resistant group and the control group, the mRNA expression of miR-21, MDR1, Bcl-2 in the inhibitor group significantly decreased (P<0.01). The mRNA expression of Bax in the inhibitor group significantly increased (P<0.001). The protein expression of P-gp, Bcl-2 in the inhibitor group significantly decreased (P<0.05). The protein expression of Bax in the inhibitor group significantly increased (P<0.001). The IC50 value in the inhibitor group significantly decreased (P<0.001). The apoptosis rate in the inhibitor group significantly increased (P<0.001). Conclusion: Silencing miR-21 can significantly increase the sensitivity of Ishikawa/DDP cells to cisplatin and promote apoptosis. The specific mechanism may be related to down regulation of MDR1, P-gp, Bcl-2, as well as up regulation of Bax.