目的 研究小白菊內(nèi)酯對(duì)惡性人腦膠質(zhì)瘤細(xì)胞系U-87 MG細(xì)胞遷移、侵襲的抑制作用, 并探討其作用機(jī)制。方法 采用CCK-8比色檢測(cè)法篩選小白菊內(nèi)酯處理細(xì)胞的IC₅₀濃度;采用細(xì)胞劃痕實(shí)驗(yàn)觀察小白菊內(nèi)酯處理0、12、48 h后U-87 MG細(xì)胞的遷移情況, 并且測(cè)量48 h后遷移的距離;采用Transwell實(shí)驗(yàn)觀察穿膜細(xì)胞數(shù), 判斷細(xì)胞的侵襲能力;并采用qPCR以及Western blotting法研究了小白菊內(nèi)酯處理后, U-87 MG細(xì)胞的SNAIL、E-Cadherin的表達(dá)變化, 以及GSK3β-Ser⁹蛋白質(zhì)磷酸化變化。結(jié)果 CCK-8比色檢測(cè)法篩選的小白菊內(nèi)酯IC₅₀濃度為39 μmol/L。與對(duì)照組比較, 在小白菊內(nèi)酯處理48 h后, U-87 MG細(xì)胞遷移的距離和穿膜細(xì)胞數(shù)都明顯減少, 且與濃度呈正相關(guān)(P<0.01);與對(duì)照組比較, 小白菊內(nèi)酯處理的U-87 MG細(xì)胞內(nèi)SNAIL基因表達(dá)下降, 同時(shí)E-Cadherin表達(dá)上調(diào), 且表達(dá)差異明顯(P<0.05);GSK3β-Ser⁹殘基磷酸化水平下降。結(jié)論 小白菊內(nèi)酯能夠抑制U-87 MG細(xì)胞的遷移、侵襲, 可能是通過下調(diào)GSK3β磷酸化從而抑制了SNAIL蛋白表達(dá)入核, 從而促進(jìn)了E-Cadherin蛋白的表達(dá)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. Methods Drug 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 39 μmol/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.

廣西壯族自治區(qū)衛(wèi)生廳自籌經(jīng)費(fèi)科研課題(Z2013624)