Inhibitory Effect Of Celatrol On Cisplatin-resistance Ovarian Cancer SKOV3/DDP Cells And Its Relevant Mechanism

ObjectiveTo investigate the inhibitory effect of Celastrol on displatin-resistance ovarian cancer SKVO3/DDP cells as well as the revelant mechanism,in order to provide effective experimental basis for clinical treatment of ovarian cancer.Methods1?The cell culture used for the investigation contains 10% fetal bovine serum,100 U,100 U/ml/ml penicillin streptomycin of RPMI1640 culture,at37? and 5% CO₂,humidity appropriate subculture in constant temperature incubator.2?We used experimental group,DDP group,Celastrol treatment group,and DDP+ Celastrol treatment group.3?MTT assay was utilized to evaluate the effect of Celastrol at various concentrations on the proliferation of SKOV3/DDP.4?We observed the effect of Celastrol on morphology of SKOV3/DDP cells under the microscope.5?We detected the cell cycle and apoptosis of SKOV3/DDP cells through the use of flow cytometry.6?The effect of Celastrol on the migration of ovarian cancer SKOV3/DDP cells were detected by cell streak test.7?By applying Transwell in vitro invasion assay,we detected the effect of varying Celastrol concentration on the invasion ability of SKOV3/DDP cells.8 ?The expression levels of proteins associated with apoptosis and invasion in SKOV3/DDP cells were determined by Western blot.Results1?MTT experiments showed that Celastrol on ovarian cancer SKOV3 /DDP cells proliferation inhibition significantly increased.?P< 0.05?.Determined by MTT assay 24 h,48 h,72 h SKOV3 and SKOV3 / DDP cells IC50,SKOV3 /DDP resistant index were 2.52,and 2.94 and 3.78.2?Compare with the control group,the number of SKOV3 / DDP cells in the experimental guoup was significantly decreased?P< 0.05?.3?With 0,1,and 1.5 mol/L Celastrol and within 24 hours,we respectively detected early apoptosis rate of SKOV3 / DDP cell at 3.42%,11.1%,and3.42%,late apoptosis rate at 4.1%,7.19%,and 4.1%,total apoptosis rate at7.52%,18.29%,and 30.9%.G1 phase cell proportion increased significantly?P< 0.05?.4?Cell streak test demonstrated respectively that 0,0.5,and 0.75 mol/L within 24 hours of cell migration were 0.0908,0.0333,and 0.0095,and within48 hours,cell migration rate were 0.2091,0.0713,and 0.0295.0.5 and 0.75mol/L including cell migration distance,Celastrol group was less than 0.5 ?Transwell invasion test respectively show that 0,0.5,0.75 mol/L Celastrol group cell membrane cell number 393± 23,241 ± 12,and 140 ± 6.0.5,and 0.75 mmol/L Celeastrol group through membrane cells were significantly fewer than the number of 0 ?mol/L celastrol group?P= 0.000 and0.000?.6 ?Western blot method detect that SKOV3/DDP cells of Cyclin A and p-AKT,NF-?B,MMP-2 and MMP-9 and P-GP protein expression levels were significantly decreased?P< 0.05?.Conclusions1?Celastrol has a significant inhibitory effect on SKOV3/DDP cells,which can affect the resistance of SKOV3/DDP cells to DDP,and promote the apoptosis of cells.2?Celatrol inhibits the DNA synthesis of tumor cells by regulating the cell cycle protein,and by reducing the Akt and NF-?B,MMP-2 and MMP-9,preventing the invasion and metastasis of tumor cells.