扶正抑瘤颗粒药物血清诱导小鼠肝癌细胞凋亡及其机制探究.docVIP

扶正抑瘤颗粒药物血清诱导小鼠肝癌细胞凋亡及其机制探究 　 作者：赵健雄，程卫东，徐瑞峰，李元青 【关键词】 ,扶正抑瘤颗粒 ［摘要］ 目的：研究中药复方扶正抑瘤颗粒药物血清对小鼠肝癌H22细胞凋亡的影响及其作用机制。方法：采用扶正抑瘤颗粒药物血清，温育体外培养的H22小鼠肝癌细胞。流式细胞仪进行细胞周期分析，检测凋亡率；透射电镜观察细胞超微结构变化；链霉亲和素生物素过氧化物酶复合物（streptavidinbiotin peroxidase complex, SABC）免疫细胞化学法观察凋亡相关蛋白Bcl2和Bax的变化。结果：扶正抑瘤颗粒药物血清可影响细胞周期，使小鼠肝癌H22细胞的增殖阻滞在G1/G0期，并可测到凋亡峰，同时可下调Bcl2蛋白的表达，上调Bax蛋白的表达（扶正抑瘤颗粒药物血清组与空白对照组比较，Plt;0.05）。结论：扶正抑瘤颗粒可通过影响细胞生长周期，调节Bcl2和Bax蛋白的表达诱导小鼠肝癌细胞凋亡。 　　［关键词］ 扶正抑瘤颗粒; 细胞凋亡; 细胞周期; 原癌基因蛋白质cbcl2; Bax 　　Effects of Fuzheng Yiliu Granulemedicated serum on apoptosis of liver cancer cells from mice and its mechanism 　　ABSTRACT Objective: To study the effects of Fuzheng Yiliu Granule (FZYLG)medicated serum on apoptosis of liver cancer cells H22 from mice and its mechanism. Methods: Liver cancer cells H22 from mice were incubated in culture media containing sera from rabbits medicated with different doses of FZYLG. Flow cytometry was used to examine the cell cycle and analyze the apoptotic rate of the H22 cells. The morphological changes of the H22 cells were observed by transmission electron microscope and the apoptosis related proteins Bcl2 and Bax were examined by streptavidinbiotin peroxidase complex (SABC) method.Results: FZYLGmedicated serum could influence the cell cycle and stop the proliferation of H22 cells at the G1/G0 phase with apoptotic peak being detected. In culture media with FZYLGmedicated sera, the expression of Bcl2 decreased while that of Bax increased as compared with that in culture medium with nonmedicated serum (Plt;0.05). Conclusion: FZYLGmedicated serum can induce apoptosis of the liver cancer cells H22 by influencing the cell cycle, downregulating the expression of Bcl2 and upregulating the expression of Bax. 　　KEY WORDS Fuzheng Yiliu Granules; apoptosis; cell cycle; protooncogene proteins cbcl2; Bax 　　抑制肿瘤细胞生长和促进其凋亡是许多抗肿瘤药物的作用机制之一。我们研制的扶正抑瘤颗粒具有扶正祛邪、化瘀解毒之功效。体内实验和临床研究发现，该药有较好的抗癌作用［1,2］，但对其抗癌机制尚缺乏深入研究。本文通过血清药理学方法，进一步观察该药对小鼠肝癌H22细胞的抑制作用，并从分子、细胞学水平探讨其抗癌机制。 　　1 材料与方法 　　1.1 实验动物 纯种新西兰家兔20只