The effects and mechanisms of Gankyrin silencing on reversing the cisplatin resistance of human ovarian cancer SKOV3/DDP cell line
10.3969/j.issn.1007-3969.2014.01.006
- VernacularTitle:Gankyrin基因沉默对人卵巢癌SKOV3/CDDP细胞顺铂耐药性的逆转作用和机制
- Author:
Qian WANG
;
Kun CHENG
- Publication Type:Journal Article
- Keywords:
Gankyrin gene;
Ovarian cancer;
Cisplatin resistant
- From:
China Oncology
2014;(1):35-40
- CountryChina
- Language:Chinese
-
Abstract:
Background and purpose: Ovarian cancer is the common gynecological cancer, and the drug resistance of anti-tumor drug was one of major reasons for therapy failure, some studies considered that there is a closed relationship between Gankyrin and drug resistance. In this study, we investigated the effects and mechanisms of Gankyrin silencing on reversing the cisplatin resistance of ovarian cancer drug-resistant SKOV3/DDP cell line. Methods:The expression of Gankyrin in SKOV3 and SKOV3/DDP cells was measured by real-time PCR assay, MTS assay was employed to determine the effect of Gankyrin on SKOV3/DDP sensitivity to cisplatin, apoptosis rate and intracellular concentration of rhodamine-123 (Rh-123) were determined by lfow cytometry, the expression of multi-drugs resistant protein MDR1, Caspase-3/8, Survivin and Bcl-2 were determined by Western blot and real-time PCR. The phosphorylation of AKT and expression of p53, NF-κB and PTEN were analyzed by Western blot assay. Results:The expression of Gankyrin was increased in SKOV3/DDP cells, Gankyrin silencing was able to increase the cisplatin sensitivity of SKOV3/DDP. Before and after gene silencing, the reverse folds (RF) to cisplatin were 1.81 and 2.45, respectively, the intracellular levels of Rh-123 were 1.73 and 2.42 fold, the apoptosis rates were 2.23 and 4.23 fold,the expressions of MDR1, Survivin and Bcl-2 were downregulated, the mRNA expressions of MDR1 were 62.8%and 21.6%, the mRNA expressions of Survivin were 24.5%and 10.3%, the mRNA expressions of Bcl-2 were 47.5%and 18.4%, the levels of Caspase-3/8, p53 and PTEN were elevated, phosphorylation of AKT and expression of NF-kB were downregulated compared with control group. Conclusion:Gankyrin silencing was able to reverse the cisplatin resistance of SKOV3/DDP cells by inhibiting the drug eflfux and promoting cell apoptosis, the PTEN/AKT/NF-κB/p53 may be the key pathway.
