The Influence Of Cinobufotalin In Vitro Effect Of Growth And Related Function Of Renal Cell Carcinoma

Objective To study the value of cinobufacini interference on 786-0 RCC cell line cultured in vitro, by observing the inhibitory effects of it on the proliferation of this cell line. To explore the potential mechanism of the cinobufacine anti-renal carcinoma activity from researching the effect of cinobufacine interference of 786-0 Renal Cancer Cell proliferation, apoptosis-inducing, invasive capacity and the effect of cell apoptosis related factors. In order to make further study of the mechanism of cinobufacine anti-renal carcinoma.Methods Cultured 786-0 Renal Cancer Cells in vitro. Used CCK8 method to observe different concentrations of cinobufacine interference 786-0 Renal Cancer Cells proliferation, to select the best inhibitory concentration group and time point. Divided 786-0 Renal Cancer Cells into three groups:control group, the best inhibitory concentration group and positive control group(5-fluorouracil), respectively effected different drugs on Renal Cancer Cells, detection of apoptosis and cell cycle of renal cell carcinoma by flow cytometry on the best inhibitory time point. Using Transwell method to detect the effect of cinobufacine on invasion of tumor cells. Using QPCR and Western blot method to detect the expression of Bcl-2、Bax and caspase-3 on RNA and protein.Results The different concentrations of cinobufacine could inhibit the proliferation of 786-0 Renal Cancer Cells, especially at the concentration of 0.1mg/ml shows the highest inhibition activity and the best inhibitory time point was 24h. The percentage of GO/G1 phase of cell cycle was increased and the percentage of G2/M phase ecreased in cinobufacin group, while there were no significant difference between the normal, control and cinobufacini group of S phase. Cinobufacin could induce cell-cycle arrest in the GO/G1 phase, inhibit DNA replication and promote the apoptosis of renal cell carcinoma cells and obviously inhibit cell invasion. The positive expression of Bax and caspase-3 genes were significantly increased, while Bcl-2 was decreased on RNA and protein level.Conclusion Cinobufacini could inhibit proliferation of human renal cell carcinoma cells with the capacity to induce cell-cycle arrest in the GO/G1 phase and inhibit DNA replication. Cinobufacini inhibit cell invasion. The mechanism of cinobufacine anti-renal carcinoma was related to inhibite expression Bcl-2, up-regulated expression Bax.