金雀异黄素与5-氟尿嘧啶单独或联合抑制人胃癌BGC-823细胞的体外研究

Combined or exclusive application of Genistein and 5-fluorouracil in inhibiting human gastric carcinoma cell line BGC-823 in vitro

摘要:

目的：研究金雀异黄素（Genistein，Gen）与5-氟尿嘧啶（5-fluorouracil，5-FU）联合应用对人胃癌细胞BGC-823的生长抑制是否具有协同作用及其可能的作用机制。方法：MTT法检测Gen、5-FU单独及联合对BGC-823细胞生长的抑制作用并用中效原理定量分析两药合用的剂量-效应关系。Gen、5-FU单独及联合处理BGC-823 细胞72 h后，流式细胞术检测细胞周期分布的变化，并在透射电镜下观察细胞超微结构的改变。结果：Gen、5-FU单用或联合应用对BGC-823细胞的生长均有显著的增殖抑制作用且呈剂量和时间依赖性。两药联合应用时，当药物效应（抑制率）大于50%时，具有协同作用[合用指数（Combina－tion index，CI）≤1]；6.06 mg/L Gen作用72 h后，95.74%的BGC-823阻滞在G2/M期；1.23 mg/L 5-FU作用72 h后，G0/G1及S期细胞分别为58.26%和41.59%，0.81 mg/L Gen与1.01 mg/L 5-FU联合应用72 h后，67.99%的BGC-823细胞阻滞在G0/G1期。Gen、5-FU单独或联合均可诱导BGC-823细胞凋亡。结论：Gen和5-FU单独或联合应用，均对BGC-823细胞具有抑制增殖的作用，在一定浓度范围内，Gen与5-FU联合应用对胃癌细胞的增殖具有协同抑制作用，Gen与5-FU可能通过阻滞细胞周期和诱导细胞凋亡而抑制胃癌细胞的生长。该研究为Gen与5-FU可能联合用于胃癌的治疗提供了依据。

Abstract:

Objective：To investigate whether combined application of Genistein（Gen） and 5-fluorouracil（5-FU） exerts synergistic ef－fect on growth inhibition of human gastric carcinoma cells BGC-823 and their possible mechanisms in vitro. Methods：Human gastric carcinoma cell line BGC-823 was exposed to Gen and 5-FU exclusively or in combination at different concentrations for 24，48 h and 72 h. 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyl tetrazolium bromide（MTT） assay was employed to investigate the inhibitory effect of Gen and 5-FU，exclusively or in combination on BGC-823 cells. Furthermore，method of median-effect principle was utilized to quantitatively analyze the dose-effect relationship of the combined effects of Gen and 5-FU. Then，BGC-823 cells were exposed to Gen and 5-FU，exclusively or in combination for 72 h，the ultramicrostructure changes of cancer cells were observed by transmission electron microscopy（TEM） and the distribution of cell cycles was detected by flow cytometry（FCM）. Results：Combined or exclusive application of Gen and 5-FU exerted significant inhibitory effect on the proliferation of BGC-823 cells in both time and dose depen－dent manner. When inhibition rate（IR） exceeding 50%，combined application of Gen and 5-FU displayed synergistic effect on growth inhibition of BGC-823 cell line. Being exposed to 6.06 mg/L Gen，1.23 mg/L 5-FU and 0.81 mg/L Gen combined with 1.01 mg/L 5-FU for 72 h，95.74% BGC-823 cells were arrested at G2/M phases，58.26% and 41.59% BGC-823 cells were arrested at G0/G1 and S phases and 67.99% BGC-823 cells were arrested at G0/G1 phases，respectively. Gen and 5-FU，used exclusively or in combination，could induce apoptosis and arrest cell cycle of BGC-823 cells. Conclusions：Combined or exclusive application of Gen and 5-FU can suppress the proliferation of BGC-823 cells in vitro. Through arresting cell cycle and inducing cell apoptosis，combined application of Gen and 5-FU can inhibit the growth of BGC-823 cells. Combined application of Gen and 5-FU exerts synergistic effect on growth of human gastric carcinoma cells within certain range of concentration. The study provides evidences for the potential combined appli－cation of Gen and 5-FU in the treatment of human gastric cancer.