摘要
目的研究小白菊内酯对恶性人脑胶质瘤细胞系U-87 MG细胞迁移、侵袭的抑制作用,并探讨其作用机制。方法采用CCK-8比色检测法筛选小白菊内酯处理细胞的IC50浓度;采用细胞划痕实验观察小白菊内酯处理0、12、48 h后U-87 MG细胞的迁移情况,并且测量48 h后迁移的距离;采用Transwell实验观察穿膜细胞数,判断细胞的侵袭能力;并采用q PCR以及Western blotting法研究了小白菊内酯处理后,U-87 MG细胞的SNAIL、E-Cadherin的表达变化,以及GSK3β-Ser9蛋白质磷酸化变化。结果 CCK-8比色检测法筛选的小白菊内酯IC50浓度为39μmol/L。与对照组比较,在小白菊内酯处理48 h后,U-87 MG细胞迁移的距离和穿膜细胞数都明显减少,且与浓度呈正相关(P<0.01);与对照组比较,小白菊内酯处理的U-87 MG细胞内SNAIL基因表达下降,同时E-Cadherin表达上调,且表达差异明显(P<0.05);GSK3β-Ser9残基磷酸化水平下降。结论小白菊内酯能够抑制U-87 MG细胞的迁移、侵袭,可能是通过下调GSK3β磷酸化从而抑制了SNAIL蛋白表达入核,从而促进了E-Cadherin蛋白的表达而引起的。
Objective To study the inhibitory effect of parthenolide on migration and invasion of malignant human brain cell lines U87 MG cell, and to explore its mechanism.MethodsDrug treatment IC50 concentration of cells was screened by CCK8 assay. Then wound healing tests were adopted to observe migration treated by parthenolide for 0, 12, and 48 h. And the migration distance was measured at 48 h. Transwell tests were used for cells penetrating observation to determine the invasive ability of the cells. Expressions of SNAIL and E-Cadherin, and GSK3β-Ser9 protein phosphorylation changes were studied by using qPCR and Western blotting methods U-87 MG cells treated by parthenolide.Results IC50 concentration of parthenolide was 39μmol/L by CCK8 assay. Compared with the control group, migration distance and the cell numbers through the membrane of U-87 MG cell were significantly reduced for 48 h after parthenolide treatment, and it was positively correlated with concentrations of the drug （P 〈 0.01）. Compared with the control group, SNAIL gene expression in U-87 MG treated by parthenolide decreased, while that of E-Cadherin increased, and there were differences （P〈 0.05）. And the levels of GSK3β-Ser9 residue phosphorylation also decreased.Conclusion Parthenolide can inhibit the migration and invasion of U-87 MG cells, which possibly caused by down-regulating GSK3β phosphorylation, preventing SNAIL protein into the nucleus, thus promoting the expression of E-Cadherin proteins.
出处
《现代药物与临床》
CAS
2015年第6期616-621,共6页
Drugs & Clinic
基金
广西壮族自治区卫生厅自筹经费科研课题(Z2013624)
