The Effect Of Cooperation Cinobufacin With 5-FU On Inhibiting Proliferation And Inducing Apoptosis Of Human Gastric Carcinoma Cells

Objective:To study the effects of cinobufacin(Cino) on inhibiting proliferation and inducing apoptosis of human gastric carcinoma cell in vitro. Besides, To study the effects the cooperation of cinobufacin with fluorouracil (5-FU) on inhibiting proliferation and inducing apoptosis of gastric carcinoma cells. Methods :The experiment was divided into control group, cinobufacin group, 5-FU group and Cino+5-FU group, Morphological changes were observed under the inverse microscope and the f luoroscope, The effect of both Cino and 5-FU on the proliferation of BGC-823 human gastric carcinoma cells was measured by means of MTT reduction assay, Ratio of apoptotic cells and cell cycle were detected by the flow cytometry. Results: gastric carcinoma cells BGC-823 presented some features of apoptosis by the inverse microscope and the fluorscope, including cell shrinkage nuclear condensation, fragmentaion, formation of apoptotic bodies. MTT assay showed that Cino could remarkably inhibit the proliferation of human gastric carcinoma BGC-823 cells in a time- and concentration-dependent manner. The rate of proliferation inhibition was more pronounced with the different dose of Cino combined with 5-FU than used individually (P<0.05), Flow cytometry analysis revealed that a typical sudiploid peak was observed before G₀/G₁ phase. Cino combined with 5-FU synergistically enhanced cell death, the rate of apoptosis is 26.5% and15. 5% separately when BGC-823 cells were treated about 48 hours Cino (lO^g/ml) or 5-FU (25[ig/ml) individually, The rate of apoptosis is 39.4% when cooperated Cino (10|j.g/ml) with 5-FU (25|a,g/ml) . The cell apoptosis was more pronounced in cells treated with both drugs than 5-FU and Cino alone(P<0. 05). Flow cytometry was used to analysze the cell cycle arrest, Cino combined with 5-FU decreased the proportion in the G0/Gi phase and increased the proportion in the S phase, and most of the cells were arrested in the S phase. Conclusion: Our results suggest that Cino has the effects of inhibiting proliferation and inducing of apoptosis in human BGC-823 gastric carcinoma cells, The effects of Cino on BGC-823 cells proliferation of inhibition were mostly in the S phase, moreover, Cino combined with 5-FU could effectively inhibit proliferation and induce apoptosis of the human carcinoma BGC-823 cells in vitro in a time- and concentration-dependent manner,...