【摘要】 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。更多还原

【Abstract】 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-803cells were treated with AKAE at different concentration.The effect on proliferation of MGC-803cells was detected by MTT assay.The flow cytometry was used to test the cell cycle of MGC-803cells.The protein and mRNA expressions of cyclin D1,p16and p21in MGC-803cells were detected by western blot and real-time PCR respectively.Results The proliferation of MGC-803cells was dose-dependently inhibited by AKAE treatment in vitro(P<0.05),and the value of IC50was(0.134±0.005)mg/ml.After treated with AKAE at the dose of 0.8mg/ml for 12h,the ratio of G1phase in MGC-803cell cycle was increased,and the ratio of S phase in MGC-803cell cycle was decreased(P<0.05).The protein and mRNA expressions of cyclin D1in MGC-803cells were obviously blocked by AKAE at the dose of(0.2-0.8)mg/ml(P<0.05).AKAE treatment could enhance the mRNA expressions of p16and p21in MGC-803cells.Compared with control group,the relative expression values of p16and p21mRNA in 0.2mg/ml group were2.30and 17.5times higher,respectively.Conclusion AKAE could inhibit the proliferation of MGC-803cells,and the mechanism is closely associated with G1-phase cell cycle arrest and expression of G1-phase related factors.更多还原