三种去甲基斑蝥素衍生物体外对人肝癌HepG-2细胞增殖的抑制作用  

Inhibitory Effect of Three Norcantharidin Derivativeson HepG-2 Cells in Vitro

作　　者：姜胜男[1] 朱欣婷[1,2] 刘云[2,3] 李三华[2,3] 雷欣睿 晏容[1,2] 李晓飞[1,2] JIANG Sheng-nan ZHU Xin-ting LIU Yun LI San-hua LEI Xin-rui YAN Rong LI Xiao-fei(Basic Medical College, Zunyi Medical University, Zunyi. Guizhou, 563000, China Guizhou Provincial College-based Key Lab for Tumor Prevention and Treatment with Distinctive Medicines, Zunyi, Guizhou, 563000, China Medical and Biological Research Center, Zunyi Medical University, Zunyi, Guizhou, 563000, China)

机构地区：[1]遵义医学院基础医学院,贵州遵义563000 [2]贵州省普通高等学校特色药物肿瘤防治重点实验室,贵州遵义563000 [3]遵义医学院医学与生物学研究中心,贵州遵义563000

出　　处：《现代生物医学进展》2017年第20期3831-3835,3841,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81260488;81660611);贵州省科技合作计划项目(黔科合LH字[2015]7516号;黔科合LH字[2015]7509号);贵州省卫计委科学基金项目(gzwjk2015-1-013);贵州省教育厅特色重点实验室建设项目(黔教合KY字[2014]212);贵州省遵义市汇川区科技计划项目(遵汇科合201630)

摘　　要：目的:探讨去甲基斑蝥素酸钠、去甲基斑蝥素酸钾、去甲基斑蝥素酸钙体外对人肝癌HepG-2细胞增殖的影响。方法:采用磺酰罗丹明(SRB法)、细胞集落形成实验、流式细胞技术检测三种去甲基斑蝥素衍生物对HepG-2细胞增殖和凋亡的影响。qRT-PCR技术考察三种去甲基斑蝥素衍生物对HepG-2细胞Bcl-2、Bax mRNA表达的影响。结果:三种去甲基斑蝥素衍生物能明显抑制HepG-2细胞的增殖,其抑制率呈剂量时效依赖性;与空白对照组相比,三种去甲基斑蝥素衍生物能显著性减少HepG-2细胞集落形成数量,并且可诱导HepG-2细胞的凋亡,凋亡率分别为8±0.56%、6.93±0.91%、6.16±0.37%;qRT-PCR显示,三种去甲基斑蝥素衍生物能抑制HepG-2细胞Bcl-2mRNA的表达,增强Bax mRNA的表达。结论:三种去甲基斑蝥素衍生物可通过下调Bcl-2 mRNA表达,上调Bax mRNA表达,诱导HepG-2细胞的凋亡,从而抑制HepG-2细胞的增殖;三种衍生物中,去甲基斑蝥素酸钠对HepG-2细胞的增殖的抑制作用最强。Objective：To explore the inhibitory effect of three norcantharidin derivatives which were named sodium norcantharidate,potassium norcantharidate and calcium norcantharidate on human hepatocellular carcinoma HepG-2 cells.Methods：The effect of three norcantharidate derivatives on HepG-2 cells proliferation was assessed with SRB assay.The cell colony forming experiment test was adopted to observe the impact of derivatives on colony formation ability of HepG-2 cells.The flow cytometry was used to examine the effect on HepG-2 cellsapoptosis,and mRNA levels of Bcl-2 and Bax were measured with RT-qPCR.Results：Three norcantharidate derivatives inhibited the proliferation of HepG-2 cells with a time and dose dependent relation.Compared with the blank control group,derivatives were able to obviously inhibit cell colony formation and induce apoptosis of HepG-2 cells.The apoptosis rate were 8± 0.56％,6.93 ± 0.91％,6.16± 0.37％,respectively.Base on the results of RT-qPCR,three norcantharidate derivatives had the ability to reduce the mRNA levels of Bcl-2 and improve the mRNA levels of Bax.Conclusion：The three kinds of norcantharidate derivatives could inhibit the proliferation and induce apoptosis of HepG-2 cells which involve in down-regulating the mRNA levels of Bcl-2 and up-regulating the mRNA levels of Bax.Among three derivatives,sodium cantharidinate shows the strongest inhibitory activity on HepG-2 cells.

关 键 词：去甲基斑蝥素酸钠 去甲基斑蝥素酸钾 去甲基斑蝥素酸钙 人肝癌HEPG-2细胞 抑制作用 

分 类 号：R-33[医药卫生] R273