姜黄素联合索拉非尼增强抑制肝癌细胞HepG-2的作用及其机制  

Study on the inhibitory effect and mechanism of Curcumin combining with sorafenib on hepatoma HepG-2 cell

作　　者：常明向[1] 吴梅梅[1] 李瀚旻[1] 

机构地区：[1]湖北中医药大学附属医院肝病研究所,湖北武汉430061

出　　处：《中西医结合肝病杂志》2016年第5期277-280,共4页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金资助项目(No.81274147)

摘　　要：目的:研究姜黄素与索拉非尼联合用药对肝癌细胞HepG-2生长的抑制作用及机制。方法:不同浓度姜黄素(10,5,2.5,1.25μg/mL),索拉非尼(5,2.5,1.25,0.625g/mL)及两者对应浓度联合应用处理肝癌HepG-2细胞不同时间后(8h,16h,24h),采用CCK-8法检测细胞增殖抑制率;姜黄素20(¨g/mL),索拉非尼10(μg/mL)及两者联合应用处理HepG-2细胞24h后,Annexin V/Pl双染法检测细胞凋亡;用流式细胞仪检测细胞周期变化情况;用Western blot法检测Caspase3、NF-Kb表达水平。结果:姜黄素、索拉非尼及联合用药对HepG-2细胞增殖均具有抑制作用,呈剂量依赖性和时间依赖性;与单药相比,姜黄素联合索拉非尼对HepG-2细胞的增殖抑制作用明显增强。姜黄素、索拉非尼均能诱导HepG-2细胞凋亡,联合用药后诱导晚期凋亡作用更强。姜黄素与索拉非尼联合用药后对HepG-2细胞周期发生明显改变,呈现出G1、G2期阻滞作用。姜黄素与索拉非尼联合用药后,下调肝癌HepG-2细胞NF-κb p65的表达,上调Caspase3的表达。结论:姜黄素与索拉非尼联合用药具有比单用姜黄素或索拉非尼更强的抗肝癌Hep-G2细胞的作用。其机制是呈现G_1、G_2期阻滞、下调肝癌HePG-2细胞NF-κb p65的表达,上调Caspase3的表达。Objective： To study the inhibitory effect and mechanism of Curcumin combining with sorafenib on HepG-2 cell. Methods： The inhibition rate of cell proliferation was determined by CCK-8 assay after HepG-2 cells were exposed to different concentrations of curcumin （ 10,5,2.5,1.25μg/mL）, sorafenib （ 5,2.5,1.25,0. 625 μg/mL） and both combination （ relative concentration ） for different time periods （ 16,24 and 48 h）. Following exposure of HepG-2 cells to curcumin （20μg/mL）, sorafenib（10μg/mL） and both combination（ relative con- centration ） for 24h,the cell apoptosis was measured by Annexin V/PI double staining,the Flowcytometry was used to detect the cell cycle , Western blot was used to detect the levels of NF-κb p65 and Caspase3. Results ： Curcumin , sorafenib and the both combination obviously in- hibited the proliferation of HepG-2 cells in a time-and concentration-dependent manner （ P 〈 0. 01 ）, the effects of combination group was more stronger. After three grug groups of treated HepG-2 ceils ,the apoptosis rates of HepG-2 cells were significantly increased , the effect of combination group showed additive effect; Compared with two single drug group, the both combination showed G1, G2 arrest ; the all NF-κb p65 expression in HepG-2 cells were down regulated, the all Caspase-3 expression were up regulated , but the both combination was more stronger. Conclusions： Curcumin combining with sorafenib has positive Anti-tumor activity on HepG-2 cell.

关 键 词：姜黄素 索拉非尼 联合用药 抗肿瘤活性 

分 类 号：R735.7[医药卫生—肿瘤]