人参皂苷Rh2调控PI3K/AKT/GSK-3β信号通路诱导人结肠癌细胞凋亡  

Ginsenoside Rh2 induced human colorectal cancer cell apoptosis through PI3K/AKT/GSK-3β pathway

作　　者：石雪萍[1] 李静[1] 冉建华[1] 熊伟[1] 李海星[1] 郭珮 陈地龙[1] 

机构地区：[1]重庆医科大学基础医学院干细胞与组织工程研究室,重庆400016

出　　处：《中国药理学通报》2017年第1期114-119,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31271368)

摘　　要：目的探究人参皂苷Rh2(ginsenoside Rh2,Rh2)诱导人结肠癌细胞SW480凋亡作用机制。方法 CCK-8法检测Rh2对SW480细胞增殖的影响;流式细胞术(Flow cyto Metry,FCM)检测Rh2对SW480细胞凋亡的影响;Hoechst33258染色观察Rh2对SW480细胞凋亡形态学的影响;Western blot检测经Rh2诱导SW480细胞中凋亡相关蛋白Bcl-2、Bax、p53、cleaved caspase-3,PI3K/AKT/GSK-3β信号通路相关蛋白PI3K、AKT、P-AKT、GSK-3β、P-GSK-3β表达量变化;LY294002、Rh2单独及联合诱导SW480细胞后,蛋白PI3K、AKT、P-AKT、GSK-3β、P-GSK-3β表达量变化。结果CCK-8结果显示Rh2呈时间浓度依赖抑制SW480细胞增殖。FCM结果显示细胞早期凋亡率由正常对照组的(0.70±0.09)%增至(11.06±1.04)%(P<0.05)。Hoechst33258结果显示,Rh2诱导SW480细胞48h后呈现典型的凋亡形态学改变。Western blot结果显示,经Rh2诱导的SW480细胞,凋亡相关蛋白Bcl-2表达降低,Bax、p53、激活型胱天蛋白酶3(cleaved caspase-3)蛋白表达增加;PI3K/AKT/GSK-3β信号通路蛋白PI3K、P-AKT、P-GSK-3β表达量与对照组比较明显减少,AKT、GSK-3β表达量无明显变化;LY294002、Rh2和LY294002与Rh2联合诱导SW480细胞后,总AKT蛋白和总GSK-3β蛋白表达量基本一致,LY294002与Rh2联合用药对SW480细胞中PI3K、P-AKT和P-GSK-3β的表达抑制作用较单独用药更加明显。结论Rh2可能是通过抑制PI3K/AKT/GSK-3β通路,激活p53信号通路,激活caspase-3,破坏Bcl-2/Bax比例,诱导结肠癌细胞SW480凋亡。Aim To investigate the effect of Ginsenoside Rh2 on apoptosis in human colorectal cancer cell SW480,and to explore the possible mechanism of it.Methods The proliferation activity of SW480 treated with Ginsenoside Rh2 was measured CCK-8 assay. Apoptosis rates were evaluated by FCM. Hoechst 33258 staining was used to observe cell nucleus morphological; change SW480 cells were treated with Ginsenoside Rh2,and the protein expressions of Bcl-2,Bax,p53,cleaved caspase-3,PI3 K,AKT,P-AKT,GSK-3β,PGSK-3β were detected by Western blot; SW480 cells were treated with LY294002,Rh2,LY294002 ＋ Rh2,the expressions of PI3 K,AKT,P-AKT,GSK-3β,PGSK-3β were detected by Western blot. Results The proliferation of SW480 cells was significantly inhibited by Ginsenoside Rh2 in dose-dependent and time-dependent manner. FCM showed the inducing apoptosis effect of Ginsenoside Rh2 was significantly different from that of control group. Hoechst 33258 staining indicated clearly cell apoptosis in Ginsenoside Rh2 treatment groups. Western blot showed Ginsenoside Rh2 decreased expression of Bcl-2,increased expression of Bax,p53 and cleaved caspase-3,PI3 K / AKT / GSK-3βpathway proteins PI3 K,P-AKT,P-GSK-3β decreased obviously,AKT and GSK-3β were not changed significantly in SW480. SW480 cells were separately treated with LY294002,Rh2,LY294002 ＋ Rh2,there were no significant difference in AKT and GSK-3β protein among all groups,and the expression of PI3 K,P-AKT,P-GSK-3β decreased more obviously in LY294002 ＋Rh2 group compared with LY294002 and Rh2 alone.Conclusion Rh2 induces colorectal cancer cell apoptosis through PI3 K / AKT / GSK-3β pathway,which activates p53 and cleaved caspase-3,and destroys the balance of Bcl-2 / Bax.

关 键 词：人参皂苷RH2 人结肠癌SW480细胞 PI3K/AKT/GSK-3β p53 Bcl-2 BAX cleaved caspase-3 凋亡 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学]