Antitumor Effect And Chemotherapy Sensitization Of Smac Mimetic On Epithelial Ovarian Cancer In Vitro

Objective In this study, IAPs was a new target for the treatment of cancer,and the Smac mimetic BV-6 was selected for this purpose.Research the antitumor effect 、chemotherapy sensitization and molecular mechanism of BV-6 on epithelial ovarian cancer in vitro.Explore the prospect of Smac mimetic as a new antitumor drug and chemosensitization agents in the clinical treatment of ovarian cancer.Methods1. MTT was used to detect the growth inhibition rate of SKOV3 cells after BV-6,determine the best effective dose and non-toxic dose.2. The appearance of apoptotic morphology of SKOV3 cells under the action of BV-6 were observed by optical microscope.3. Using RT-PCR 、Western blot to detect the expression of TNF-α m RNA and proteins in SKOV3 cells under the best effective dose of BV-6.4. MTT was used to detect the growth inhibition rate of SKOV3 cells under the non-toxic dose of BV-6 combination DDP, observe cisplatin sensitivity by the changes of DDP IC50.5. Using RT-PCR、Western blot to detect the expression of Caspase m RNA and proteins in SKOV3 cells under the non-toxic dose of BV-6 combination DDP.Results1. MTT results: BV-6 can effectively inhibit the growth of SKOV3 cells in vitro,and increase with time and dose.Then determine the best effective dose of0.1umol/L,non-toxic dose of 0.01umol/L.The non-toxic dose growth inhibition rates of 24h、48h 、72h were(2.26±0.21)%、(4.50±0.38)%、(8.22±0.66)%,and efective dose group were(33.88 ±0.94)%、(48.67 ±0.87)%、(58.40±0.79)%.Effective dose significantly inhibited tumor cell growth, and induced cell apoptosis.2. RT-PCR results: the effective dose group 24 h 、 48 h 、 72 h TNF-α m RNA expression gradually increased, the increasing multiple is 0.60 、 1.19 、 1.68 compared with the control group. There was no significant difference in the expression of TNF-α m RNA between 24 h and 48 h, and the other groups had statistical difference(P < 0.05).3. Western blot results: the effective dose group at 24h、48h、72h TNF-α protein expression was significantly higher,the increasing multiple is 0.23、0.78、1.89 compared with the control group, differences between groups has statistical significance(P < 0.05).4. MTT showed that 24h、48h、72h DDP IC50 were 14.42ug/ml、6.73ug/ml、3.19ug/ml,combined with non-toxic BV-6,the DDP IC50 were 10.19ug/ml 、3.53ug/ml、1.32ug/ml, they are significantly reduced. The SKOV3 cell growth inhibition rate non-toxic BV-6 group, DDP group and double medicine group were(4.50±0.38)% 、(47.95±0.98)% 、(60.78±0.66)% in 48 h, combined with non-toxic BV-6 significantly enhanced the cell growth inhibitory effect of DDP(P < 0.05).5. RT-PCR and Western blot showed that there was no significant difference in the expression of Caspase m RNA and protein with control group, while the expression of double medicine groups was significantly higher than DDP group,and the difference was statistically significant(P < 0.05).Conclusions1. High-dose BV-6 inducing apoptosis of SKOV3 cells to play antitumor effect,and in the effective dose range was time concentration dependent.2. Low-dose BV-6 can increase the sensitivity of SKOV3 cells to cisplatin, and non-toxic dose did not cause additional damage to cells.3. BV-6 cause activation of caspase, promote the expression of TNF-α, stimulate tumor cell mitochondria apoptosis pathway and death receptor pathway of apoptosis.