黄芩素通过诱导活性氧产生抑制MET/Akt信号通路促进人结肠癌SW480细胞凋亡  

Baicalein promotes human colon cancer cell SW480 apoptosis by inhibiting MET/Akt signaling pathway via reactive oxygen species generation

作　　者：徐佳丽 路上云 王佳 石东星 邱服斌 XU Jia-li;LU Shang-yun;WANG Jia;SHI Dong-xing;QIU Fu-bin(Department of Nutrition and Food Hygiene,School of Public Health,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学公共卫生学院营养与食品卫生学教研室,山西太原030001

出　　处：《中国药理学与毒理学杂志》2022年第11期812-818,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：山西省基础研究计划项目(202103021223215);山西省博士启动基金项目(SD2016);山西医科大学博士启动基金(XD2025)。

摘　　要：目的 研究黄芩素对结肠癌细胞SW480凋亡的诱导作用及其机制。方法 人结肠癌细胞SW480经黄芩素0,20,40和80μmol·L^(-1)处理48 h后,通过结晶紫染色法测定细胞形态和存活率;试剂盒检测活性氧(ROS)水平、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性;流式细胞术检测细胞凋亡;Western印迹法检测活化胱天蛋白酶3和活化聚腺苷二磷酸核糖聚合酶(PARP)蛋白表达水平及酪氨酸激酶受体(MET)蛋白、蛋白激酶B(Akt)和重组人源组蛋白H3蛋白(H3)磷酸化水平。结果 结晶紫染色结果显示,黄芩素可浓度依赖性抑制SW480细胞存活(P<0.01,r=-0.928)。ROS及抗氧化酶检测结果显示,与细胞对照组相比,黄芩素20,40和80μmol·L^(-1)组ROS生成明显增加,而SOD和CAT活性则明显降低(P<0.05,P<0.01)。流式细胞术结果显示,与细胞对照组相比,黄芩素40和80μmol·L^(-1)组细胞凋亡率显著升高(P<0.01)。Western印迹结果表明,与细胞对照组相比,黄芩素处理后SW480细胞中活化胱天蛋白酶3和活化PARP表达上调(P<0.05,P<0.01),MET,Akt和H3磷酸化水平下调(P<0.05)。结论 黄芩素通过诱导ROS产生抑制MET/Akt信号通路诱导结肠癌SW480细胞凋亡。OBJECTIVE To investigate the inhibitory effect and mechanism of baicalein(Be) on colon cancer cell line SW480. METHODS After treatment with baicalein 0, 20, 40 and 80 μmol·L^(-1)for 48 h, cell proliferation was determined by crystal violet staining. Kits were used to detect levels of reactive oxygen species(ROS), superoxide dismutase(SOD) and catalase(CAT). The apoptosis was detected by flow cytometry. Western blotting was used to detect the expression levels of apoptotic related proteins cleaved-caspase 3, cleaved-poly(ADP-ribose) polymerase(PARP), protein phosphorylation levels of mesenchymal-epithelial transition factor(MET), protein kinase B(Akt), and histone H3(H3).RESULTS Crystal violet staining showed that baicalein could inhibit cell proliferation in a concentrationdependent manner(P<0.01, r=-0.928). The results of flow cytometry showed that the apoptosis rate of baicalein 40 and 80 μmol · L^(-1)groups was significantly higher than that of the cell control group(P<0.01). Compared with the cell control group, the expressions of cleaved-caspase 3 and cleaved-PARP were up-regulated(P<0.05), protein phosphorylation levels of MET, Akt and H3 were down-regulated(P<0.05), fluorescence intensity of ROS was increased, while SOD and CAT activities were decreased(P<0.05) in baicalein treatment groups. CONCLUSION Baicalein induces apoptosis of colon cancer cell line SW480 by inhibiting MET/Akt signaling pathway via ROS induction.

关 键 词：黄芩素 结肠癌 细胞凋亡 活性氧 SW480细胞 

分 类 号：R285.5[医药卫生—中药学] R966[医药卫生—中医学]