Objective: To investigate the synergistic effects on cell apoptosis and growing restriction of SKOV3 cells by the combination of compound herbal medicinal prescription (CHMP) with cisplatin (DDP).
Methods: Cisplatin and two CHMP for tonic quality(CHMP1) and activating blood circulation (CHMP2), which was medicated serum, were prepared and used to treat the human ovarian carcinoma cell line SKOV3. By serum pharmacologic method, the growth and apoptosis of SKOV3 cell were observed at different time points(24,48,72, 96 h) with different concentrations of medicated serum. Coefficient of drug interaction (CDI) between CHMP, and CHMP2 was studied by MTT method. The effects of control group(A group),CHMP1 group(B group),CHMP2 group (C group), DDP group(D group), CHMP1 + DDP group(E group), CHMP2 + DDP group(Fgroup)to SKOV3 cell were studied by flow cytometry; and the cell apoptosis was observed by agarose electrophoresis; the expressions of TNFR1, caspase-8 on each group were analyzed by Western blot method.
Results: Synergistic effects were found between herbal medicinal mixtures and DDP, Restraining rate of SKOV3 and CHMP serum concentration was not in a dose-dependent manner as DDP was. CDI between CHMPI and CHMPS was found to be significant difference (CDI of CHMP1, CHMP2 and DDPwas 0.66, 0.58 respectively). It showed that the combined treatment was able to get better effect than single drug treatment. The performed agarose electrophoresis revealed the extracted DNA to show a typical ladder patterns for cell apoptosis. The analysis results of western blot showed the increased expressions of TNFR1 and caspase-8 after combined using of medicine, which were accord to the rates of apoptosis.
Conclusions: CHMP drug granules show the synergistic effects with DDP, and the suppressing functions in the course of cell proliferation, and the inducing effect on apoptosis of human ovarian carcinoma cell line SKOV3 in vitro. And this mechanism is showed to be sponsored by the activation of TNFR1 and Caspase-8.