Down-regulation of miR-205-5p sensitizes HNE1/DDP to cisplatin induced apoptosis in vitro
10.16438/j.0513-4870.2018-1142
- VernacularTitle:下调miR-205-5p增强鼻咽癌细胞HNE1/DDP对顺铂诱导凋亡的敏感性
- Author:
Wei-man ZHAO
1
;
Zhi-yong DONG
1
;
Zong-fen SHI
1
;
Xing-yue LU
1
;
Hao LIU
1
;
Pei ZHANG
1
Author Information
1. School of Pharmacy, Bengbu Medical College, Bengbu 233030, China
- Publication Type:Research Article
- Keywords:
nasopharyngeal carcinoma cell;
cisplatin;
miR-205-5p;
rug resistance;
apoptosis
- From:
Acta Pharmaceutica Sinica
2019;54(7):1200-1206
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to investigate the effect of down-regulation of miR-205-5p by transfection of miR-205-5p inhibitor on the sensitivity of HNE1/DDP cells to cisplatin (DDP) induced apoptosis and explore the underlying mechanism. qRT-PCR was used to detect the expression of miR-205-5p in HNE1 or HNE1/DDP cells. The expression level of miR-205-5p was analyzed after transfecting HNE1/DDP cells with miR-205-5p inhibitor. MTT assay was used to evaluate the inhibitory effect of DDP alone or in combination with miR-205-5p inhibitor on the proliferation of HNE1/DDP or HNE1 cells. Apoptosis of cells treated with miR-205-5p inhibitor alone or in combination with DDP (8 μmol·L-1) was assessed using flow cytometry with PI staining, with the nucleus was counterstained with DAPI staining. The expression of Bax, Bak, Mcl-1, or Bcl-2 was analyzed by Western blot. HNE1/DDP cells showed a high level of expression of miR-205-5p, and the expression of miR-205-5p was significantly decreased by transfection of miR-205-5p inhibitor. Down-regulation of miR-205-5p significantly increased the sensitivity of HNE1/DDP cells to DDP (P<0.05). Transfection of miR-205-5p inhibitor enhanced the sensitivity of HNE1/DDP cells to DDP induced apoptosis. Treatment of HNE1/DDP cells with miR-205-5p inhibitor combined with DDP (8 μmol·L-1) for 24 h resulted in an apoptotic rate of 28.93% ± 2.50%, significantly higher than that treated with miR-205-5p inhibitor (9.83% ± 1.31%) or DDP alone (10.83% ± 1.70%) (P<0.05). DAPI staining showed that HNE1/DDP cell nucleus became significantly condensed and fragmented in miR-205-5p inhibitor combined with DDP group. The combined group up-regulated the expression of Bax and down-regulated the expression of Bcl-2 in HNE1/DDP cells. Therefore, down-regulation of miR-205-5p can enhance the sensitivity of HNE1/DDP cells to cisplatin induced apoptosis, and the mechanism may involve up-regulation of Bax and down-regulation of Bcl-2 expression.
