| Objective This study aims to investigate the effect of antioxidant pyrrolidine dithiocarbamate (PDTC) on cell growth, NF-κB activity ,and the expression of VEGF and IL-8 in the ovarian carcinoma OVCAR3 cell line, in order to evaluate PDTC as a potential drug for chemoprevention and chemotherapy of ovarian carcimoma.Methods 1. The cytotoxity of PDTC, DDP, or PDTC combined with DDP to OVCAR3 cell line were detected by MTT method. 2. The apoptotic rate and Sub-G1 peak was analyzed by FCM using PI staining method. 3.EMSA (gel mobility shift assay):The constitutive nuclear NF-kappa B activity in OVCAR3 cells as well as 0VCAR3 cells treated by PDTC (1μM ,5μM or 10μM) was measured by EMSA. And the NF-kappa B activity were also measured by EMSA in negative control group, IL-1β stimulation group(10ng/ml,1h),and PDTC inhibition groups( pretreated with 1μM and 10μM PDTC for 30 minutes before stimulated by 10ng/ml IL-1β for 1h).4. The expression of IL-8 and VEGF mRNA in OVCAR3 cells was detected by RT-PCR after the cells were treated by IL-1β or PDTC for certain time.Results 1.PDTC could significantly inhibit the proliferation of OVCAR3 cells in a dose-dependent and time-dependent manner. 2.Administration of DDP combined with PDTC significantly improved cytotoxity of DDP to ovarian cancer cells. The inhibition rate was only 3.1% when cells were treated with 0.01 μg/ml DDP alone for 24h. The inhibition rate increased to 19.0% when cells were pretreated with 1μM PDTC before exposed to 0.01μg/ml DDP. The differences between both methods were significant at different concentrations of DDP, P<0.05. 3.PDTC could induce apoptosis of human ovarian cancer OVCAR3 cells. The apoptotic rate was 2.29% andthe cell apoptosis peak (CAP) was not obvious in 10 u M PDTC group. The apoptosis rate was 38.5% and obvious subdiploid CAP in 100 y M PDTC. There was significant differences between the two groups with different concentration of PDTC, P<0.05.4.OVCAR3 cells expressed constitutive NF- k B DNA binding activity. PDTC significantly decreased this constitutive level while the activation of IL-1 3 resulted in a remarkable increase of its activity. PDTC significantly decreased activated level of NF- k B induced by IL-1 3 in a dose-dependent manner. There was significant difference between inhibition group and stimulation group, P<0.05.5.IL-1 ￡ promoted the expression of VEGF and IL-8 mRNA in OVCAR3 cells and the mRNA level reached to peak after IL-1 P administered for 4 hours. The activated mRNA level of VEGF and IL-8 induced by IL-1 P was significantly inhibited by pretreatment of PDTC and there was significant difference between PDTC inhibition group and IL-1 P stimulation group, P<0.05.Conclusion1. PDTC could inhibit the proliferation and induce apoptosis of OVCAR3 cells so as to promote anti-cancer effect of DDP.2. PDTC could significantly and effectively decrease constitutive and inductive NF-kB DNA binding activity of OVCAR3 cells. PDTC can be used as NF-kB inhibitor in research on ovarian carcinoma.3. The expression of VEGF and IL-8 mRNA induced by IL-1 P stimulation was effectively inhibited by PDTC.4. Antioxidant PDTC might provide novel prospect for chemoprevention and chemotherapy of ovarian carcinoma.
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