Objective To explored the role of autophagy activation in paclitaxel(PTX)-resistant ovarian cancer cells and the effects of autophagy inhibition on the treatment of resistant cancer cells.Methods The ovarian cancer cell lines SKOV3 and PTX-resistant ovarian cancer cells were used.MTT assay was used to determine the cell viability.Apoptosis rates were detected by Annexin V/PI staining assay.The protein expression was determined by western blot(WB).The cells were transfected by LC3-GFP-RFP using lipofectamine 2000 and the fluorescence was detected by laser scanning confocal microscope.Results The inhib-itory effect of PTX on SKOV3/PTX cells was significantly reduced compared with SKOV3 cells and inhibitory concentration50%(IC50)of PTX on SKOV3 cells and SKOV3/PTX cells were 41.91±2.92 μM and 114.6±17.6 μM,respectively.The apoptot-ic effect of PTX on SKOV3/PTX cells was significantly reduced.It was proved that SKOV3/PTX cells showed increased autophagy activation after LC3-GFP-RFP overexpression through detecting the expression of LC3 and P62.Autophagy of SKOV3/PTX cells was inhibited after treatment with autophagy inhibitors,chloroquine(CQ)and 3-MA and the inhibitory effect and apoptosis-indu-cing ability of PTX on SKOV3/PTX cells was improved.Conclusion Autophagy activation has a protective role in the PTX-re-sistant ovarian cancer cells.Inhibition of autophagy enhanced the cytotoxic effect of PTX-resistant ovarian cancer cells.
关键词
卵巢癌/紫杉醇耐药细胞/紫杉醇/自噬/SKOV3
Key words
Ovarian cancer/PTX-resistant cancer cells/PTX/Autophagy/SKOV3
维普
万方数据