GMF-β与MEK信号通路cross-talk介导榄香烯的抗胶质瘤作用

Cross-talk between GMF-β and MEK signaling pathway mediates the inhibitory effect of elemene on glioblastoma

目的探讨神经胶质成熟因子-β(Glia Maturation Factor-β,GMF-β)在榄香烯致人U87MG胶质瘤细胞增殖抑制中的作用。方法以不同浓度榄香烯作用于人U87MG胶质瘤细胞,噻唑蓝(Methyl Thiazolyl Tet-razolium,MTT)法检测细胞活性。免疫沉淀联合Western blot法检测经榄香烯处理的U87MG细胞中总GMF-β及磷酸化GMF-β(p-GMF-β)蛋白的表达水平。通过RNA干扰技术使GMF-β的表达沉默,然后以MTT法检测榄香烯的抗胶质瘤增殖能力。Western blot法检测榄香烯对丝裂原活化蛋白激酶激酶3/6(Mitogen-activated Protein Ki-nase Kinase 3/6,MEK3/6)的激活作用是否因GMF-β的沉默而受到影响。结果榄香烯显著抑制了人U87MG细胞的增殖,并可使GMF-β、MEK3和MEK6的磷酸化水平上调。沉默GMF-β的表达导致MEK3和MEK6的磷酸化水平下调,榄香烯的抗胶质瘤细胞增殖作用减弱。结论 GMF-β与MEK信号通路的交互作用(cross-talk)介导了榄香烯的抗人胶质瘤细胞增殖作用。

Objective To investigate the role of Glia Maturation Factor-β（GMF-β）in the anti-glioblastoma proliferation effect of elemene. Methods Human U87MG glioblastoma cells were treated with elemene at different drug doses.Cell viability was determined using a Methyl Thiazolyl Tetrazolium（MTT）assay.The levels of total GMF-β and p-GMF-β in elemene-treated U87MG cells were examined by immunoprecipitation and Western blot analysis.Upon GMF-β silencing using RNA interference,the antitumor action of elemene was evaluated in a MTT assay,and Mitogen-activated Protein Kinase Kinase 3/6（MEK3/6）and p-MEK3/6 expressions were examined by semiquantitative Western blot analysis. Results Elemene inhibited the proliferation activity of U87MG glioblastoma cells and increased phosphorylation levels of GMF-β,MEK3 and MEK6.Conversely,the expression of silencing GMF-β reversed the anti-glioblastoma effect of elemene and impaired the phosphorylation of MEK3/6. Conclusion The cross-talk between GMF-β and MEK signaling pathway underlays the inhibitory effect of elemene on the proliferation of human glioblastoma cells.