Down-regulating osteopontin expression by shRNA interference enhances chemotherapy sensitivity of cervical cancer hela cells
10.3724/SP.J.1008.2013.01299
- Author:
Fang FANG
1
Author Information
1. Department of Obstetrics and Gynecology, Changhai Hospital, Second Military Medical University
- Publication Type:Journal Article
- Keywords:
Apoptosis regulatory proteins;
Osteopontin;
Rna interference;
Ulterine cervical neoplasms
- From:
Academic Journal of Second Military Medical University
2013;34(12):1299-1303
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of silencing osteopontin (OPN) expression by short-hairpin RNA (shRNA) interference on chemotherapy sensitivity of cervical cancer HeLa cells and the possible mechanism. Methods HeLa cells were transfected with eukaryotic expression vector pGCsi3. 0 carrying OPN shRNA via liposome (shOPN group). Untransfected HeLa cells (Con group) and those transfected with empty plasmids (shNon group) served as controls. HeLa cells in all the groupswere treated with different concentrations of cisplatin (0, 0. 5, 1, and 2 (μg/mL) and paclitaxel (0, 50, 100, and 500 nmol/L) for 24 h, respectively; the apoptosis in Hela cells was analyzed by flow cytometry. The expressions of apoptosis related protein cleaved caspase-3, Bct-xL, Bct-2, and Bax were examined by Western blotting analysis. Results The apoptotic rate in shOPN group ([44. 53±2. 78]%) was significantly higher than those in shNon group ([15. 34±2. 18]%) and Con group ([15. 37± 1. 03]%) after treatment with 2 jug/mL cisplatin (P<0. 05); and significant difference was also found between the apoptotic rates after treatment with 500 nmol/L paclitaxd (shOPN group: [51. 46±1. 49]%, shNon group: [19. 16 ± 1. 87] %, Congroup: [17. 03±2. 37]%; P<0. 05). Down-regulating OPN expression significantly enhanced the cisplatin-induced activation of cleaved caspase-3 (P<0. 01), and it also resulted in inhibition of Bd^/Bd-xL. expression and up- regulation of Bax expression. Conclusion OPN silencing can sensitize cervical cancerHeLa cells to chemotherapeutic agents by promoting HeLa cells apoptosis. RNA interference mediated depletion of OPN may be a promising strategy for the new adjuvant chemotherapy treatment of cervical cancer.
