雪灵芝水提物对人胃癌MGC-803细胞增殖及细胞周期的影响

Effects of Arenaria Kansuensis Aqueous Extract on Proliferation and Cell Cycle of Human Gastric Cancer Cell Line MGC-803

目的探讨雪灵芝水溶性提取物（arenaria kansuensis aqueous extract，AKAE）对人胃癌细胞株MGC-803细胞增殖及细胞周期的影响。方法体外培养的MGC-803细胞经不同浓度AKAE处理，采用MTT法检测受试细胞增殖水平；应用流式细胞术检测AKAE处理12h后各组细胞的细胞周期；Western blot检测不同浓度、不同时间AKAE处理组MGC-803细胞中cyclin D1蛋白表达水平；实时定量PCR检测各组MGC-803细胞中cyclin D1、p16和p21基因的mRNA相对表达量。结果AKAE对体外培养的MGC-803细胞增殖具有浓度依赖性抑制作用（P〈0．05），IC50为（0．134±0．005）mg／ml；经AKAE处理12h，0．8mg／ml组MGC-803细胞的G1期细胞比例高于对照组、S期细胞比例低于对照组（P〈0．05）；（0．2～0．8）mg／ml浓度的AKAE处理，对MGC-803细胞中cyclin D1的蛋白及mRNA表达具有明显抑制作用（P〈0．05）；AKAE促进MGC-803细胞中p16、p21基因mRNA表达，0．2mg／ml处理组p16、p21mRNA相对表达量分别为对照组的3．3倍和18．5倍。结论雪灵芝水提物（AKAE）对MGC-803细胞增殖具有抑制作用，其机制与G1期阻滞和对G1期相关因子表达的影响有关。

Objective To investigate the effects of arenaria kansuensis aqueous extract （AKAE） on proliferation and cell cycle of human gastric cancer cell line MGC-803. Methods MGC-803 cells were treated with AKAE at different concentration. The effect on proliferation of MGC-803 cells was detected by MTT assay. The flow cytometry was used to test the cell cycle of MCK2-B03 cells. The protein and mRNA expressions of cyclin D1, p16 and p21 in MGC-803 cells were detected by western blot and real-time PCR respectively. Results The proliferation of MGC-803 cells was dose-dependently inhibited by AKAE treatment in vitro （P〈0. 05）, and the value of IC50 was （0. 134 ＋ 0. 005）mg/ml. After treated with AKAE at the dose of 0. 8 mg/ml for 12 h, the ratio of G1 phase in MCA2-803 cell cycle was increased, and the ratio of S phase in MGC-803 cell cycle was decreased（P〈0. 05）. The protein and mRNA expressions of cyclin D1 in MGC-803 cells were obviously blocked by AKAE at the dose of （0. 2-0. 8） mg/ml（P% 0. 05）. AKAE treatment could enhance the rnRNA expressions of p16 and p21 in MGC-803 cells. Compared with control group, the relative expression values of p16 and p21 mRNA in 0. 2 mg/ml group were 2. 30 and 17. 5 times higher, respectively. Conclusion AKAE could inhibit the proliferation of MGC-803 cells, and the mechanism is closely associated with G1-phase cell cycle arrest and expression of Gt-phase related factors.