摘要
目的研究黄芩素(BAI)对胃癌细胞系MGC-803细胞自噬的影响。方法使用(0、 5、 15、 25、 50)μmol/L BAI处理胃癌MGC-803细胞24、 48、 72 h,噻唑蓝(MTT)法检测细胞的增殖活性,吖啶橙(AO)染色结合免疫荧光细胞化学染色观察微管相关蛋白1轻链3(LC3)和P62的表达确定MGC-803细胞的自噬情况, Western blot法检测细胞LC3、 P62、磷脂酰肌醇3激酶(PI3K)、磷酸化的PI3K(p-PI3K)、蛋白激酶B(AKT)、 p-AKT的蛋白水平。结果与对照组相比, BAI可显著抑制MGC-803细胞的增殖,并呈时间和剂量依赖性;BAI处理的MGC-803细胞内酸性溶酶体显著增加, LC3表达增强;BAI处理后可显著降低PI3K和AKT蛋白的磷酸化, LC3-Ⅱ/LC3-Ⅰ比值增加、上调P62蛋白的表达。结论黄芩素抑制PI3K/AKT信号通路诱导MGC-803细胞发生自噬。
Objective To investigate the effect of baicalein(BAI) on autophagy in gastric cancer cell line MGC-803. Methods MGC-803 cells were treated with 0, 5, 15, 25, 50 μmol/L BAI for 24, 48, 72 hours. The proliferation activity of MGC-803 cells was detected by MTT assay. Acridine orange(AO) staining combined with immunofluorescence cytochemical staining was performed to observe the expression of microtubule-associated protein 1 light chain 3(LC3) and P62 to determine autophagy in MGC-803 cells. The protein levels of LC3, P62, phosphatidylinositol 3-kinase(PI3K), phosphorylated PI3K(p-PI3K), AKT, and p-AKT were detected by Western blot analysis. Results Compared with the control group, BAI significantly inhibited the proliferation of MGC-803 cells in a time-and dose-dependent manner. BAI-treated MGC-803 cells showed a significant increase in acid lysosomes and increased LC3 expression. BAI treatment significantly decreased phosphorylation of PI3K and AKT proteins, increased the ratio of LC3-Ⅱ/LC3-Ⅰ and up-regulated the expression of P62 protein. Conclusion Baicalein could inhibit PI3K/AKT signaling pathway and induce autophagy in MGC-803 cells.
作者
乔丹
李英顺
邢健
孙沛林
王莹
张莹莹
陈丽艳
任香善
林贞花
金京春
朴英实
QIAO Dan;LI Yingshun;XING Jian;SUN Peilin;WANG Ying;ZHANG Yingying;CHEN Liyan;REN Xiangshan;LIN Zhenhua;JIN Jingchun;PIAO Yingshi(Cancer Research Center,Yanbian University,Yanji 133002;Key Laboratory of Science and Technology Department of Jilin Province,Yanji 133002;Medical College,Yanbian University,Yanji 133002;Hongqi Hospital Affiliated to Mudanjiang Medical College,Mudanjiang 157011;Hospital Affiliated to Yanbian University,Yanji 133000,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2019年第7期613-618,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81860651)
