柴胡皂甙d对肝癌SMMC-7721细胞COX-2表达的-第三军医大学学报.DOC

基金项目：国家自然科学基金资助 基金项目：国家自然科学基金资助项目(No 作者简介 HYPERLINK mailto: ：和水祥，电话：（029 E-mail: HYPERLINK mailto:hesx123@126.c hesx123@126.com 和水祥1，朱占芳1,2，卢新兰1，赵 刚3，陈静宏4，王治伦4，罗金燕3 （710061西安,西安交通大学：第一附属医院消化内科1，医学院地方性骨病研究所4 ，710049西安，西安交通大学校医院2，710004西安，西安交通大学第二附属医院消化内科3） [摘要] 目的: 观察柴胡皂甙d(SSd)对肝癌SMMC-7721细胞血管内皮生成因子（VEGF）和血管生成素-2（Ang-2）表达的影响，探讨其可能的抗癌作用机制。方法: 肝癌SMMC-7721细胞株经LPS诱导，加入不同终浓度的SSd（2.5，5，10，20 mg·L-1）作用48h，并以Nimesulide作为对照，采用MTT法检测细胞增殖，免疫细胞化学法测定肝癌细细内VEGF和Ang-2蛋白表达，RT-PCR法检测VEGF-mRNA表达，ELISA法检测培养上清中VEGF含量。结果: SSd可显著抑制肝癌SMMC-7721细胞的生长、增殖。SSd作用组肝癌细胞VEGF蛋白、mRNA表达和培养上清中VEGF含量水平均显著降低，Ang-2蛋白表达亦显著降低，与Nimesulide组变化相似（P﹥0.05）。在2.5～10mg·L-1作用终浓度范围内，SSd对培养上清中VEGF含量影响呈现一定的量效关系。结论: SSd可能通过COX-2 /VEGF和Ang-2信号通路，发挥其抗癌作用。 [关键词] 肝肿瘤；肿瘤细胞，培养的；柴胡皂甙；血管内皮生长因子；血管生成素-2； [中图分类号] R735.7 [文献标识码] A Effects of Saikosaponins-d on VEGF and Ang-2 expression of human hepatocellular carcinoma cell line SMMC-7721 He Shuixiang1, Zhu Zhanfang1，2, Lu Xinlan1, Zhao Gang3, Chen Jinghong4, Wang Zhilun4, Luo Jinyan3.( 1Department of Gastroenterology, First Affiliated Hospital, Medical School of Xi’an Jiaotong University, 4Institute of Endemic Bone Disease，College of Medicine, Xian, Shanxi Province 710061 2Hospital of Xi’an Jiaotong University，Xian, Shanxi Province, 710049 3Department of Gastroenterology, Second Affiliated Hospital, Medical School of Xi’an Jiaotong University，Xian, Shanxi Province,710004, China) [Abstract] Objective To observe influence of saikosaponins-d on the expression of VEGF and Ang-2 in hepatocellular carcinoma cells, and explore the underlying mechanism of anti-tumor by saikosaponins-d. Methods The human hepatocellular carcinoma cell line SMMC-7721 was cultured in vitro and induced by LPS then exposed to nimesulide, and several concentrations(2.5，5，10，20 mg·L-1) of saikosaponins-d for 48 hours, the cell proliferative activity was measured by MTT method，the protein expression of VEGF and Ang-2 were assayed using immunocytochemistry, the mRNA expressi