四嗪二甲酰胺诱导人胃癌AGS、SGC7901细胞系凋亡及其分子机制的实验研究  

Study on apoptosis of human gastric carcinoma cell line AGS and SGC7901 induced by N,N'-di-(m-methylphenyl)3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine1,4-dicarboamide and its molecular mechanism

作　　者：吕亚萍[1] 周永列[2] 胡惟孝[1] 邱莲女[2] 饶国武[1] 王文松[2] 

机构地区：[1]浙江工业大学药学院,杭州310014 [2]浙江省人民医院中心实验室,杭州310014

出　　处：《药物分析杂志》2008年第7期1105-1113,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：浙江省科技厅资助项目(2003c33019)

摘　　要：目的:观察四嗪二甲酰胺(ZGDHu-1)体外抑制胃癌细胞株 AGS 和 SGC7901增殖并诱导细胞凋亡及其作用机制。方法:将不同浓度的 ZGDHu-1与 AGS 和 SGC7901细胞在体外培养,用台盼蓝染色、SRB 法、5′-溴-2′脱氧尿苷(Brdu)-ELISA 法观察 ZGDHu-1对 AGS 和 SGC7901细胞增殖的抑制作用;用细胞形态学、DNA 凝胶电泳、DNA 含量及细胞周期分析、Annexin-V/PI 双标记、Hoechst33258荧光染色等技术检测细胞凋亡。用流式细胞术观察经 ZGDHu-1作用后 AGS 和SGC7901细胞 bcl-2、bax、P53蛋白质和 P38MAPK 和 Stat3磷酸化表达的变化的表达改变。结果:ZGDHu-1能抑制 AGS和 SGC7901细胞增殖和活力,呈现作用时间和剂量的量效关系。AGS 和 SGC7901细胞与 ZGDHu-1作用后,大部分细胞阻滞于 G_2-M 期;出现典型的细胞肜态改变,DNA 片断化,亚 G1峰检出并显著增加,Annexin-V+/PI-表达升高,Ho-echst33258荧光染色后出现凋亡细胞的特征性改变等均证实 ZGDHu-1能诱导 AGS 和 SGC7901细胞凋亡。AGS 和SGC7901细胞经不同浓度的 ZGDHu-1体外培养后,bcl-2蛋白有所下调,但主要是上调 bax 和 P53蛋白,导致 bax/bcl-2比值明显增高,均并呈现剂量依赖性;磷酸化 P38MAPK 表达增强,而磷酸化 Stat3表达降低。结论:ZGDHu-1通过诱导细胞凋亡抑制 AGS 和 SGC7901细胞增殖,其机制可能与上调 bax 和 P53的表达,增强 P38MAPK 的磷酸化和抑制 STAT3的活化有关。Objective：To investigate the possibility of human gastric carcinoma cells apoptosis induced by N,N′ - di - （m - methylphenyl） -3,6 - dimethyl - 1,4 - dihydro - 1,2,4,5 - tetrazine - 1,4 - dicarboamide in vitro and its molecular mechanism Method： Different concentration of ZGDHu - 1 and different times of cultivate were used to treat AGS and SGC7901 cell. The proliferation inhibition was analysed by alive cell count, SRB assay and Brdu - ELISA. Cell apoptosis was analysed by cell morphology, DNA agarose gel electrophoresis, DNA content, Annexin - V/PI and Hoechst 33258 labeling method. The expressions of bcl - 2, bax, P53 and phosphorylated p38MAPK and STAT3 were analysed by flow cytometry. Results：ZGDHu -1 can inhibit AGS and SGC7901 cell proliferation and viability within a certain range of treating time and dose, and a majority of AGS and SGC7901 cells were arrested in G2 -M phase. The AGS and SGC7901 cell apoptosis was confirmed by type cell morphology, DNA fragment, sub -G1 phase, Hoechst33258 and Annexin/PI Labeling method with a time and dose related manner. The expression of bax,p53 and bax/bcl -2 ratio and phosphor -p38MAPK protein was significantly increased and phosphor- STAT3 down - regulated by the treatment of ZGDHu - 1. Conclusion： ZGDHu - 1 can significantly inhibited AGS and SGC7901 proliferation by inducing tumor cell apoptosis in vitro. The mechanism may associate with its up - regulation of bax,P53 and phosphor - p38MAPK and down - regulation of phosphor - STAT3 during the apoptosis process.

关 键 词：四嗪二甲酰胺 胃癌 细胞株 AGS和SGC7901 增殖 细胞凋亡 

分 类 号：R917[医药卫生—药物分析学]