IGFBP-5及p53与汉防己甲素抑制乳腺癌MCF-7细胞增殖的关系研究  

Study on relationship between IGFBP-5/p53 axis and anti-proliferation effect of tetrandrine in MCF-7 cells

作　　者：周林云[1,2] 任文艳 廖云鹏[1,2] 王涵[1,2] 朱茄慧 胡莹[1,2] 李福书 周娅 何百成 

机构地区：[1]重庆医科大学药理教研室,重庆400016 [2]重庆医科大学重庆市生物化学与分子药理学重点实验室,重庆400016

出　　处：《中国药理学通报》2018年第1期38-43,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81372120)

摘　　要：目的研究汉防己甲素(tetrandrine,Tet)抑制乳腺癌MCF-7细胞增殖的作用及可能的分子机制。方法采用CCK-8、流式及Western blot分析不同浓度Tet对MCF-7细胞增殖与凋亡的影响;通过real-time PCR、Western blot检测Tet对MCF-7细胞IGFBP-5表达的影响;应用CCK-8分析Tet及IGFBP-5过表达或沉默表达对Tet抑制乳腺癌MCF-7细胞增殖作用的影响;利用Western blot检测Tet对MCF-7细胞p53表达的影响,以及过表达IGFBP-5和(或)Tet对乳腺癌MCF-7细胞p53表达的影响。结果 Tet呈浓度和时间依赖性抑制MCF-7细胞增殖,诱导G1期阻滞,并降低PCNA表达水平;同时,Tet明显增加MCF-7细胞凋亡比例,升高Bad的蛋白水平,降低Bcl-2的蛋白水平。Tet上调MCF-7细胞IGFBP-5的表达水平,过表达IGFBP-5增加Tet对MCF-7细胞的增殖抑制作用,而沉默IGFBP-5则减弱Tet的这种作用。Tet升高MCF-7细胞的p53蛋白水平,降低MDM2的蛋白水平;过表达IGFBP-5增强Tet对p53表达的促进和对MDM2表达的抑制作用。结论 Tet对MCF-7细胞增殖具有抑制作用,机制可能与其上调IGFBP-5而增强p53信号的功能有关。Aim To study the effect of tetrandrine( Tet) on proliferation of MCF-7 breast cancer cells and the possible mechanism underlying this biological process. Methods CCK-8, flow cytometric and Western blot were introduced to analyze the effect of Tet on proliferation and apoptosis in MCF-7 cells. Real-time PCR and/or Western blot assay were employed to detect the effect of Tet on expression of IGFBP-5,p53 and MDM2. CCK-8 and recombinant adenovirus were utilized to determine the effect of IGFBP-5 on the proliferation inhibitory effect of Tet. Western blot assay was introduced to evaluate the effect of IGFBP-5 on p53 which was induced by Tet. Results Tet inhibited the proliferation,arrested cell cycle at G1 phase and decreased the expression of PCNA concentration dependently in MCF-7 cells. Meanwhile,Tet increased the percentage of apoptotic cells,the level of Bad and reduced the level of Bcl-2. Tet increased the expression of IGFBP-5 either mRNA or protein,over-expression of IGFBP-5 enhanced the anti-proliferation activity of Tet in MCF-7 cells,but knockdown of IGFBP-5 attenuated this effect of Tet. Tet increased the level of p53 and decreased that of MDM2,and exogenous IGFBP-5 enhanced the effect of Tet on p53 and MDM2,respectively. Conclusion Tet can inhibit the proliferation of MCF-7 cells,and this activity is partly mediated by increasing the function of p53 signal,which may be triggered by the Tet-induced IGFBP-5.

关 键 词：汉防己甲素 乳腺癌 MCF-7 增殖 IGFBP-5 p53信号 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]