Down-regulation of pannexin 2 channel enhances cisplatin-induced apoptosis in testicular cancer I-10 cells.
10.12122/j.issn.1673-4254.2020.08.04
- Author:
Yanxue YAO
1
;
Shuying DONG
1
;
Chenlu ZHU
1
;
Miao HU
1
;
Baolong DU
1
;
Xuhui TONG
1
Author Information
1. School of Pharmacy, Bengbu Medical College, Bengbu 233030, China.
- Publication Type:Journal Article
- Keywords:
apoptosis;
cisplatinum;
pannexin 2;
testicular cancer
- MeSH:
Antineoplastic Agents;
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Cisplatin;
Connexins;
Down-Regulation;
Drug Resistance, Neoplasm;
Humans;
Male;
Testicular Neoplasms
- From:
Journal of Southern Medical University
2020;40(8):1090-1096
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of down-regulation of pannexin 2 (Panx-2) channels on cisplatin-induced apoptosis in I-10 cells.
METHODS:The expression of Panx-2 protein in testicular cancer cells was detected with Western blotting. The testicular cancer cell line I-10 was transfected with two short hairpin RNA (shRNA1 and shRNA2) Lipofectamine, the empty vector (NC group) or Lipofectamine2000 (blank control group), and the changes in the expression of Panx-2 was detected with Western blotting. The effects of transfection with a Panx-2 inhibitor on surviving fraction of the cells treated with cisplatin (16 μmol/L) for 24 h, 48 h and 72 h was assessed with MTT assay, and the clonogenic capacity of the cells was evaluated with colony-forming assay. At 8 h after incubation with 16 μmol/L cisplatin, AnnexinV/PI double staining was used to detect the early apoptosis of the cells. After 24 h of treatment with 16 μmol/L cisplatin, the cells were examined for expressions of caspase-3, Bcl-2 and Bax using Western blotting.
RESULTS:The expression of Panx-2 was significantly increased in cisplatin-resistant I-10/DDP ( < 0.001) cells and Tcam-2/DDP ( < 0.01) cells as compared with I-10 cells and Tcam-2 cells. Transfection of I-10 cells with shRNA1 and shRNA2 resulted in significantly decreased Panx-2 expression ( < 0.05) and significantly reduced cell surviving fraction ( < 0.001). In the presence of cisplatin, the cells in NC group showed a higher clonogenic efficiency than those in shRNA1 and shRNA2 groups ( < 0.001). The early-stage apoptosis rate of the cells in shRNA1 and shRNA2 groups were significantly higher than that in NC group ( < 0.01). Panx-2 knockdown in I-10 cells significantly increased caspase-3 and Bax expressions ( < 0.05) and significantly decreased the expression of Bcl-2 ( < 0.01).
CONCLUSIONS:Down-regulation of Panx-2 channel enhances cisplatin-induced apoptosis in cultured testicular cancer cells.
