橘红素对人胃癌AGS细胞自噬的作用及其机制研究  

Role of tangeretin on autophagy in human gastric cancer AGS cells and its mechanism

作　　者：陈圣敏 龚永昌 隋璐 楚敏 董杨 CHEN Sheng-min;GONG Yong-chang;SUI Lu;CHU Min;DONG Yang(Experimental Teaching Centre,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学教学实验中心

出　　处：《中国药理学通报》2019年第12期1671-1676,共6页Chinese Pharmacological Bulletin

基　　金：上海市教委重点学科资助项目(No J50301)

摘　　要：目的探讨橘红素(Tangeretin,Tan)对人胃癌AGS细胞自噬的作用及其机制。方法Western blot检测Tan对LC3II、p62及p-Akt蛋白表达的影响,以及自噬抑制剂CQ和促进剂Rapa干预后,LC3II和p62蛋白表达的变化。Annexin V/PI双染,流式细胞术检测CQ对Tan诱导细胞凋亡作用的影响。结果作用24 h,Tan可呈剂量依赖地增强LC3II蛋白(P<0.05,P<0.01)和p62蛋白(P<0.05)的表达;作用48 h,Tan可呈剂量依赖地增强p62蛋白的表达(P<0.05,P<0.01)。作用24 h和48 h,与Tan单独处理组比较,CQ与Tan联合处理可明显增强LC3Ⅱ和p62蛋白的表达(P<0.01),而Rapa和Tan联合处理则下调LC3Ⅱ和p62蛋白的表达(P<0.05)。10μmol·L^-1 CQ与60μmol·L^-1 Tan联合作用48 h,CQ+Tan组与Tan组比较,细胞凋亡率明显增加(P<0.01)。Tan作用24 h和48 h,p-Akt蛋白的表达均呈剂量依赖地下降(P<0.05,P<0.01)。结论橘红素可以抑制人胃癌AGS细胞的自噬流,造成自噬体的堆积,促进AGS细胞凋亡,Akt蛋白的磷酸化可能参与其中。Aim To investigate the effect of tangeretin on autophagy of human gastric cancer AGS cells and its molecular mechanisms. Methods Western blot was used to detect the effect of tangeretin on expression of LC3, p62 and Akt phosphorylation, and the effects of autophagy inhibitor CQ and activator Rapa on tangeretin-regulated autophagy. Annexin V/PI double staining was used to quantitatively detect the effect of CQ on apoptosis of cells induced by tangeretin. Results 24 h after treatment,tangeretin enhanced the expression of LC3 Ⅱ protein( P < 0. 05,P < 0. 01) and p62( P <0. 05) in a dose-dependent manner. 48 h after treatment,tangeretin enhanced the expression of p62 protein in a dose-dependent manner( P < 0. 05,P <0. 01),and had no significant effect on expression of LC3 Ⅱ protein. At 24 h and 48 h,the expression of LC3 Ⅱ and p62 increased significantly in CQ + Tan group compared with that of Tan group( P < 0. 01).Compared with Tan group,the expression of LC3 Ⅱ and p62 protein decreased in Rapa + Tan group,and there was a statistical difference( P < 0. 05). The apoptotic rate of AGS cells in CQ + Tan group was significantly higher than that in Tan group( P < 0. 01). The phosphorylation levels of Akt protein were reduced by tangerin for 24 and 48 h in a dose-dependent manner( P< 0. 05,P < 0. 01). Conclusions Tangeretin may promote cell apoptosis by inhibiting the autophagic flux in human gastric cancer AGS cells. Akt phosphorylation might be involved in the autophagy and apoptosis.

关 键 词：橘红素 胃癌 自噬 自噬流 凋亡 AKT 

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