Research On The Reversal Of Cisplatin-Resistance In Human Ovarian Carcinoma By Neferin In Vitro

Objective To detect the effect of neferine (NEF) on the proliferation, on the sensitivity of cisplatin (CDDP) and the expression of GST-Ï€gene in ovarian cancer cell line SKOV3/DDP. To investigate preliminary the reverse role of NEF on cisplatin-resistant human ovarian carcinoma cell line.Methods (1) Human ovarian cancer cell line SKOV3 and its cisplatin-resistant cell line SKOV3/DDP was cultured to the logarithmic growth phase in vitro; (2) The expression of GST-Ï€was examined by immunohistochemical method in human ovarian cancer cells; (3) To screen non-toxic doses of NEF on drug-resistant cell line by MTT(Methyl thiazoly tetrazolium) assay, and The changes of DDP resistance was determined after cells were treated with nontoxic dose of NEF by MTT assay; (4) The expression level of GST-Ï€gene was detected by semi-quantitative RT-PCR.Results (1) NEF has cell growth inhibitory roll on SKOV3/DDP, and the inhibitory rate of proliferation corresponding rise with the increase in the concentration,it is a role of drug dose-dependent, 1,5,10 ug / ml NEF have not a significant cytotoxicity (cell inhibition rate less than 10%) to SKOV3/DDP, The non-toxic doses of 10ug / ml is as a reversal of doses in this experiment; when the DDP combined with NEF of the non-toxic doses , the DDP group 's IC50 decreased on SKOV3/DDP cell, and the synergistic interaction on SKOV3/DDP cells increased with time-dependent, respectively, 1.15, 1. 91, 2.28 folds (P<0.01);(2)the result of immunocytochemistry shows, in the SKOV3/DDP group,the expression of GST-Ï€was higher than SKOV3 group under the microscope, when the tumer cells were treated by NEF after 72h ,the expression of GST-Ï€was decreased, color lighter; (3) RT-PCR results showed : Compared with the two groups,the expression level of GST-Ï€mRNA was significantly inhibited in the SKOV3/DDP+NEF group(P < 0 . 01).Conclusion it is the one of important reasons that over-expressed GST-Ï€gene promote chemotherapy resistance in ovarian cancer, NEF can reduced over-expression of GST-Ï€on drug-resistant cell SKOV3/DDP and effectively reverse the resistance of ovarian cancer to cisplatin and increase their drug sensitivity.