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目的:通过研究红花多糖(safflower polysaccharide,SPS)对人肝癌SMMC-7721细胞增殖的影响,探讨SPS抗肿瘤作用的分子机制。方法:体外培养人肝癌SMMC-7721细胞,加入含不同质量浓度SPS(0,0.02,0.04,0.08,0.16,0.32,0.64,1.28 g·L-1)的培养液,分别培养24,48 h,用四甲基偶氮唑盐比色法(MTT法)检测SPS对细胞增殖的影响;用免疫组化法检测0.16,0.32,0.64 g·L-1SPS作用肝癌细胞48 h,细胞的细胞周期素B1(cyclin B1)蛋白表达;采用实时荧光定量PCR技术(realtime PCR,RT-PCR)和蛋白免疫印迹(Western blot)法检测细胞分裂周期25B(Cdc25B)基因的表达。结果:肝癌细胞的生存率随SPS浓度和作用时间的增加而降低,而1.28 g·L-1SPS组除外。SPS作用48 h后SMMC-7721细胞的cyclin B1蛋白、Cdc25B蛋白和mRNA表达降低,并具有剂量依赖性。结论:SPS可能通过抑制细胞周期相关基因的表达,诱导肝癌细胞增殖阻滞,进而起到抗肿瘤作用。
Objective: To investigate the effect of safflower polysaccharide (SPS) on the proliferation of human hepatocellular carcinoma SMMC-7721 cells and explore the molecular mechanism of antitumor effect of SPS. Methods: Human hepatocellular carcinoma SMMC-7721 cells were cultured in vitro, and the medium containing different concentrations of SPS (0,0.02,0.04,0.08,0.16,0.32,0.64,1.28 g · L -1) was added and cultured for 24,48 h, The effect of SPS on cell proliferation was detected by MTT colorimetric assay. The expression of cyclin was detected by immunohistochemistry B1 (cyclin B1) protein expression was detected by real-time PCR (Real-time PCR, RT-PCR) and Western blotting method to detect the expression of cell cycle 25B (Cdc25B). Results: The survival rate of hepatocellular carcinoma cells decreased with the increase of SPS concentration and time, except for 1.28 g · L-1SPS group. The expression of cyclin B1 protein and Cdc25B protein and mRNA in SMMC-7721 cells decreased in a dose-dependent manner after SPS for 48 h. Conclusion: SPS may inhibit the proliferation of hepatocellular carcinoma cells by inhibiting the expression of cell cycle-related genes and thus may play an antitumor effect.