Establishment of a cisplatin-induced multidrug resistance cell line SK-Hep1/DDP
- VernacularTitle:人肝癌顺铂耐药细胞系SK-Hepl/CDDP的建立
- Author:
Zhou YUAN
1
;
Ling XIAN-LONG
Author Information
1. 第三军医大学附属新桥医院
- Keywords:
Hepatoma;
cisplatin;
multidrug resistance;
SK-Hep1 cell line;
MDR1;
MRP1;
MRP2
- From:Chinese Journal of Cancer
2010;29(2):176-181
- CountryChina
- Language:Chinese
-
Abstract:
Background and Objective:Multidrug resistance(MDR)is a major obstacle in the chemotherapy of cancer patients.The aim of this study was to establish a mutidrug-resistant cell line SK-Hep1/DDP and explore its molecular mechanism of the MDR. Methods: SK-Hep1/DDP cell line was induced by pulse treatment using a high concentration of cisplatin(DDP)in vitro.The chemoresistance indexes of cells were evaluated bv CCK-8 assays.The protein of MDRl(ABCB1),MRPl(ABCC1),MRP2(ABCC2)and Bax were detected by Western blotting, and the effect of MDR1 inhibitor cyclosporine A(CsA)on expression of MDRl proteins in SK-Hep1 and SK.Hep 1/DDP cell lines. Flow cytometry was performed to determine the distribution of the cell cycle and cell apoptosis ralio.Results:The SK-Hep1/ DDP cells were 13.76 times more resistant to DDP in comparison with SK.Hep1 cells.and SK-Hep1/DDP cells also exhibited cross-resistance to many other chemotherapeutic agents(for example adramycin and 5-fuorouracil).MDRl,MRPl and MRP2 protein expressions were significantly higher in the SK-Hep1/DDP than in the SK-Hep1(P<0.01),but Bax was lower in the SK-Hep1/DDP than in the SK-Hep1(P<0.01).There was no obvious influence between SK-Hep1 and SK-Hep1/DDP cells in the expression of MDRl bv MDR1 inhibtor CsA(P>0.05).The percentages of cells in G_2/M and S phase were significantly increased in SK-Hep1/DDP in comparison with those in SK-Hep1[(20.67±5.69)%vs.(12.14士3.36)%;(42.20±2.65)%vs.(27.91±2.16)%;P<0.01].After the cells were exposed to 10 μg/mL DDP for 24 h.the cell apoptosis rate of SK-Hep1/DDP was decreased in comparison with SK-Hep1,but it was increased in those with pretreatment of MDRl inhibitor CsA as compared with those without pretreatment.Conclusions: A reliable multidrug-resistant human hepatoma cell line SK-Hep1/DDP is successfully established.The MDR mechanisms of this cell lines are closely relaled to the over-expression of MDR1, MRP1 and MRP2,lower expression of Bax, and the attenuated cell apoptosis induced by chemotherapeutic agents.
