维甲酸Ro 40-8757对人肿瘤细胞系的作用及其机制探讨  

Effects of Retinoid Ro 40-8757 on Human Cancer Cell Lines In Vitro

作　　者：吕晶[1] 宋今丹[1] 

机构地区：[1]卫生部细胞生物学重点实验室中国医科大学分子生物学研究所,辽宁沈阳110001

出　　处：《癌症》2002年第10期1051-1056,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金重大项目(No.3904005)

摘　　要：背景与目的:维甲酸类化合物对多种恶性肿瘤具有诱导分化、抑制增殖、诱导凋亡等作用。本文旨在研究一种新合成的维甲酸Ro40-8757对4种体外培养的人肿瘤细胞系生长增殖、形态结构及细胞周期的影响,并探讨其机制。方法:用MTT法测定Ro40-8757对4种人肿瘤细胞系(人结肠癌细胞系CCL-187和CCL-229、人胰腺癌细胞系JF-305和AsPC-1)的增殖抑制作用;用光镜和电镜观察其对敏感细胞系CCL-187细胞形态的影响;用流式细胞术研究其对细胞周期的影响;最后,用Westernblot法检测在给药前后CCL-187细胞周期蛋白p16、p21和p27的变化情况,以探讨其可能的机制。结果:Ro40-8757对4种细胞系均有显著的生长抑制作用,并具有时间、浓度依赖性;显微镜观察未见细胞有明显的细胞毒性、凋亡和分化特征性改变;药物作用后4种肿瘤细胞系细胞周期阻滞于G0/G1期;Ro40-8757作用12h后CCL-187细胞系p21和p27同时升高,24、48h逐渐下降,72h又再次升高144h升至最高;p16在加药前后始终阴性表达。结论:Ro40-8757对人结肠癌细胞系CCL-187和CCL-229、人胰腺癌细胞系JF-305和AsPC-1有明显的增殖抑制作用,并呈时间、浓度依赖性;这种抑制作用是通过将细胞周期阻滞于G0/G1期发挥作用的,细胞周期蛋白p21和p27在其中起作用引起细胞周期阻滞。Background & Objectives: The retinoids are potent anti tumor drugs affecting cellular proliferation and inducing cell differetiation or apoptosis. This study was designed to investigate the anti proliferative effect of a new developed retinoid, Ro 40 8757, on cell structure, cell cycle and cell cycle proteins of four human cancer cell lines in vitro in order to reveal its probable mechanism. Methods: MTT assay was used to determine the anti proliferative effects of Ro 40 8757 on human cancer cell lines CCL 187,CCL 229, JF 305, and ASPC 1. Microscopy was used to observe CCL 187 morphological changes. Flow cytometry was performed to investigate the influence of Ro 40 8757 on cell cycle. Western blot analysis was conducted to detect the cell cycle proteins p16, p21, and p27 as to discuss the possible mechanism. Results: Ro 40 8757 significantly inhibited the growth of the four cancer cell lines in a dose , time dependent manner and without any signs of cytotoxicity, differentiation, or apoptosis. After treating with Ro 40 8757, the cell cycle of CCL 187 was arrested at G0/G1 phase. Both p21 and p27 were rapidly increasing at the first 12 hours after exposed to the agent, then decreasing slowly in the 24, 48 hours, and increasing again in 72 to 144 hours. P16 did not express at all either before or after agent treatment. Conclusion: The results suggest that Ro 40 8757 inhibits the growth of human cancer cell lines in vitro by means of cell cycle arrest (mediated by up regulating cell cycle protein P21 and P27) instead of cytotoxic effects, differentiation, or apoptosis.

关 键 词：维甲酸 Ro40-8757 肿瘤 增殖抑制 细胞周期 MTT法 结肠癌 胰腺癌 

分 类 号：R730.53[医药卫生—肿瘤]