DBDCT致HL02细胞毒性的初步作用机制研究  

Study on the toxic mechanism of DBDCT on HL02 cells

作　　者：王晓燕[1] 唐莉[1] 李云兰[1] 李青山[1] 

机构地区：[1]山西医科大学药学院,山西太原030001

出　　处：《毒理学杂志》2016年第4期256-259,264,共5页Journal of Toxicology

基　　金：国家自然科学基金(30973603);山西省研究生优秀创新项目(20113070);山西医科大学博士启动基金(03201319)

摘　　要：目的研究新型有机锡化合物DBDCT对肝细胞HL02的毒性作用及其机制。方法 MTT法测定DBDCT对HL02细胞活力的影响,计算IC50,并于光学显微镜下观察细胞形态的改变;紫外分光光度法测定LDH的释放量;流式细胞术测定细胞凋亡与细胞周期;Western blot测定周期蛋白Cdc2的表达。结果 DBDCT可抑制HL02细胞的增殖,且随着药物浓度的增加,细胞形态逐渐模糊、细胞固缩变圆,表现出明显的细胞毒性;24 h IC50为5.77μmol/L;DBDCT可引起LDH的大量释放,并将细胞阻滞于G2/M期,细胞凋亡率增加,且呈现明显的量效关系;细胞周期蛋白依赖性激酶Cdc2的表达则随着给药浓度的增加而降低。结论 DBDCT可通过改变细胞膜的通透性,降低Cdc2的表达,引起G2/M期细胞阻滞与细胞凋亡,进而产生明显的肝细胞毒性。Objective To study the toxic mechanism of new diorganotin（ IV） compound DBDCT on liver cell line HL02. Methods Cytotoxicity of DBDCT on HL02 cells was assessed by MTT method,and the IC50 value was calculated. Meanwhile,the morphological changes were examined under optical microscope. The level of lactate dehydrogenase（ LDH） was determined using ultraviolet spectrophotometric method. Cell apoptosis and cell cycle were analyzed by flow cytometry. The expression of Cdc2 was detected by Western blotting. Results The cell growth was inhibited in a dose dependent manner,and the IC50 value was 5. 77 μmol / L at 24 h. Cell morphology became gradually blurred,atrophic,presenting obvious cell toxicity. The level of LDH was increased significantly,and the cell was arrested at G2 / M phase followed with cell apoptosis in a dose dependent manner. Cyclin dependent kinase Cdc2 was down-regulated following exposure to DBDCT. Conclusions DBDCT resulted in the obvious liver cell toxicity via reducing the expression of Cdc2,changing the permeability of the cell membrane,and causing the G2 / M phase arrest and cell apoptosis.

关 键 词：DBDCT HL02细胞 毒性机制 CDC2 

分 类 号：R996[医药卫生—毒理学]