黄芪皂苷Ⅱ增加5-氟尿嘧啶对人肝癌细胞株HepG2增殖抑制作用  

Increasing inhibition of human hepatic cancer HepG2 cells to 5-fluorouracil by AstragalosideⅡ

作　　者：武超[1] 许杜娟[1,2] 杨翠[1] 夏泉[1,2] 张医浩 

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学第一附属医院药剂科,合肥230022

出　　处：《安徽医科大学学报》2016年第1期78-82,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:11040606M222)

摘　　要：目的探讨黄芪皂苷Ⅱ增加5-氟尿嘧啶对人肝癌细胞株(Hep G2)增殖抑制作用及其可能机制。方法采用MTT方法检测5-氟尿嘧啶单独或联合黄芪皂苷Ⅱ作用于细胞后,5-氟尿嘧啶对Hep G2的抑制率;Hoechst33342染色检测细胞凋亡;Western blot检测丝裂原活化蛋白激酶(ERK1/2)、磷酸化丝裂原活化蛋白激酶(p-ERK)以及凋亡蛋白多聚ADP-核糖聚合酶(PARP)、活化凋亡蛋白多聚ADP-核糖聚合酶(Cleaved PARP)、活化半胱氨酸蛋白酶蛋白-3(Cleaved caspase 3)、活化半胱氨酸蛋白酶蛋白-9(Cleaved caspase 9)的表达。结果 MTT法结果显示,5-氟尿嘧啶呈浓度和时间依赖性抑制肝癌Hep G2细胞增殖,联用黄芪皂苷Ⅱ可以显著增加5-氟尿嘧啶对Hep G2细胞增殖抑制作用(P<0.05);Western blot法结果显示,联合使用黄芪皂苷Ⅱ和5-氟尿嘧啶能增加Cleaved PARP表达(P<0.05);Hoechst33342验证联合用药组较5-氟尿嘧啶组细胞凋亡水平升高;此外,Western blot结果还显示,黄芪皂苷Ⅱ抑制p-ERK1/2蛋白表达;联用ERK1/2特异性抑制剂PD98059明显增加5-氟尿嘧啶诱导的细胞凋亡率以及凋亡蛋白Cleaved caspase3、Cleaved caspase9表达(P<0.05)。阻断ERK1/2后,黄芪皂苷Ⅱ促进5-氟尿嘧啶诱导的凋亡蛋白Cleaved caspase3、Cleaved caspase9表达以及细胞凋亡水平并未明显增加。结论黄芪皂苷Ⅱ增加5-氟尿嘧啶对人肝癌细胞株Hep G2增殖抑制作用,其机制可能与抑制p-ERK1/2的表达有关。Objective To investigate the Astragaloside II increasing inhibilion of human hepatic cancer （HepG2） cells by 5-fluorouracil and its possible mechanism. Methods The HepG2 inhibiting rate by 5-fluorouracil after a- lone or jointly acting on cells was detected by MTT method ; cells apoptosis was detected by Hoechst33342 staining. The expressions of ERK1/2,p-ERK1/2,PARP, Cleaved PARP, Cleaved caspase 3 and Cleaved caspase 9 proteins were measured by Western blot. Results MTT staining showed that 5-fluorouracil inhibited the proliferation of HepG2 cells and presented dose and time-dependence. Furthermore, combining the Astragaloside II with 5-fluorou- racil could further significantly enhance the effect of 5-fluorouracil on cancer cells inhibition （ P 〈 0.05 ）. Western blot result proved that combining the Astragaloside Ⅱ with 5-fluorouracil could increase the expression of Cleaved PARP （P 〈 0. 05 ）. Hoechst33342 staining result further validated the level of apoptosis cells was far more in- creased in the combination group than the 5-fluorouracil group. Moreover, Western blot result also showed that As- tragaloside Ⅱ inhibited the phosphorylation of ERK1/2. Combining the inhibitor of ERK1/2 （PD98059） with 5-flu- orouracil may increase apoptosis rate and the expressions of apoptosis related proteins like Cleaved caspase3, Cleaved easpase 9 in HepG2 ceils. Astragaloside Ⅱ showed negative increasing trend of apoptosis rate and the ex- pressions of apoptosis related proteins like Cleaved caspase 3, Cleaved easpase 9 by 5-fluorouracil after blocking ERK1/2 in HepG2 cells. Conclusion Astragaloside Ⅱ could increase inhibition of human hepatic cancer HepG2 cells by 5-fluorouraeil, and the mechanism may be related to inhibiting p-ERK1/2 pathway.

关 键 词：黄芪皂苷Ⅱ 5-氟尿嘧啶 HEPG2 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]