丹参酮IIA对低氧条件下人肝癌HepG2细胞增殖、凋亡的影响及与HIF-1α、VEGF和野生型P53蛋白表达的关系  

Effects of tanshinone IIA on proliferation,apoptosis and expression of HIF-1α,VEGF and wild-type P53 in human hepatoma Hep G2 cells under hypoxia

作　　者：刘丽璇 吴灵飞[1] 邓巍[1] 周小涛[1] 陈锐沛 项梦琦 郭益添[1] 蒲泽锦[1] 李国平 

机构地区：[1]汕头大学医学院第二附属医院消化内科,广东汕头515041

出　　处：《中国病理生理杂志》2014年第12期2155-2160,共6页Chinese Journal of Pathophysiology

基　　金：广东省中医药局资助项目(No.20121165)

摘　　要：目的:探讨低氧条件下丹参酮IIA(Tan IIA)对人肝癌Hep G2细胞增殖和凋亡的影响及其分子机制。方法:用氯化钴(Co Cl2)创建低氧模型,实验分为常氧对照组、低氧对照组和低氧+Tan IIA处理组。不同浓度的Tan IIA分别作用于低氧下人肝癌Hep G2细胞24 h、48 h和72 h,采用MTT法测定Tan IIA对低氧下Hep G2细胞增殖的抑制作用。不同浓度的Tan IIA分别作用于低氧条件下Hep G2细胞24 h和48 h后,Hoechst 33258染色法检测细胞核的形态学变化并计算凋亡率。不同浓度的Tan IIA作用于低氧条件下人肝癌Hep G2细胞48 h后,Western blotting检测低氧诱导因子1α(HIF-1α)、血管内皮生长因子(VEGF)和野生型P53的蛋白表达情况。结果:低氧条件下Tan IIA以时间和剂量依赖方式抑制Hep G2细胞的生长和增殖。Tan IIA作用于低氧下的Hep G2细胞后,可见典型的凋亡细胞形态学特征,细胞凋亡率呈时间、剂量依赖性的增加。Western blotting免疫印迹法显示常氧对照组的HIF-1α和VEGF表达较低,而低氧对照组的HIF-1α、VEGF蛋白表达较常氧组升高,低氧下随着Tan IIA浓度的升高,HIF-1α和VEGF蛋白的表达明显降低,野生型P53蛋白的表达随着Tan IIA浓度的升高而升高。结论:低氧条件下,Tan IIA能抑制肝癌Hep G2细胞增殖并诱导其凋亡,其机制可能与抑制HIF-1α和VEGF蛋白表达,上调P53蛋白表达有关。AIM ： To investigate the effects of tanshinone IIA （Tan IIA） on proliferation, apoptosis and its mo- lecular mechanism in human hepatoma HepG2 cells under hypoxic condition. METHODS ： Hypoxia model was established by treatment with cobalt chloride （ CoCt2 ）. The cells were divided into normoxia control group, hypoxia control group and hypoxia combined at different concentrations of Tan IIA groups. After HepG2 cells were incubated with different concentra- tions of Tan IIA （0. 5, 1.0, 2. 0, 5.0 and 10. 0 mg/L） for24 h, 48 h and 72 h under hypoxic condition, the cell prolifer- ation was determined by MTY assay. After Tan IIA was added to the media at different concentrations for 24 h and 48 h, the apoptotic cells were observed by Hoechst 33258 staining. The protein levels of hypoxia-inducible factor 1 alpha （ HIF- lot）, vascular endothelial growth factor （VEGF） and wild-type P53 were detected by Western blotting after cultured with different concentrations of Tan IIA for 48 h. RESULTS ： Tan IIA inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner. Tan IIA induced the typical morphology of apoptotic ceils and increased the apoptotic rate in a dose- and time-dependent manner after treatment with 1.0 mg/L - 5.0 mg/L for 24 h and 48 h under hypoxic condition. The protein levels of HIF-lct and VEGF were weakly expressed in HepG2 ceils under normoxia but up-regulated after incu- bated under hypoxia for 48 h. The protein expression of HIF-lot and VEGF were decreased with the increase in the concen- tration of Tan IIA under hypoxia. The protein expression of wild-type P53 was increased with the increase in the concentra- tions of Tan IIA under hypoxia. CONCLUSION ： Tan IIA significantly inhibits the proliferation and induces the apoptosis of human hepatoma HepG2 cells under hypoxia, which may be related to the down-regulation of HIF-la and VEGF and up- regulation of wild-type P53.

关 键 词：丹参酮IIA 低氧 HEP G2细胞 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]