摘要
目的探讨姜黄素(CCM)通过抑制Hedgehog信号通路抑制γ-射线治疗脑胶质瘤诱发的上皮间质样转化(EMT)。方法以人脑胶质母细胞系LN229和U251为研究对象,设置空白对照组、CCM处理组、γ-射线处理组和联合处理组。采用CCK-8增殖抑制实验检测γ-射线处理、γ-射线联合姜黄素处理对脑胶质瘤细胞系增殖抑制的影响; Western blot实验检测脑胶质瘤细胞系中Hedgehog通路激活状态以及EMT转化状态; Transwell侵袭小室实验检测CCM和γ-射线对肿瘤细胞侵袭及迁移能力的影响。结果在CCK-8增殖抑制实验中,LN229细胞系增殖抑制的状况与γ-射线呈剂量依赖性和时间依赖性,但在U251细胞系中该抑制作用不明显; Western blot显示,经γ-射线放射治疗后,在两个细胞系中均发现E-cadherin和Claudin表达水平下降,β-catenin和Vimentin的表达水平上升;经γ-射线处理96 h、CCM处理24 h后,两个细胞系Hedgehog信号通路激活水平受抑制,同时EMT转化也受到抑制; Transwell侵袭小室实验进一步显示,在γ-射线处理96 h、CCM处理24 h后,两个细胞系的侵袭和迁移能力均下降。结论姜黄素通过抑制Hedgehog信号通路,进一步抑制γ-射线处理脑胶质瘤细胞系后诱发的EMT转化,从而提高γ-射线对杀伤胶质瘤细胞的效力。
Objective To explore the inhibition of curcumin on epithelial-mesenchymal transition(EMT) caused by γ-irradiation(γ-IR) via suppressing the Hedgehog signaling pathway.Methods LN229 and U251 cell lines were divided into four groups respectively,the negative control group,CCM group,γ-IR group and CCM + γCIR group.CCK-8 assay was used for the inhibition of proliferation caused by γ-irradiation.Western blot assay was used to detect the activity of Hedgehog signaling pathway and EMT status;Transwell assay was used to measure the effect of curcumin and γ-irradiation on the invasion and migration capacity of the glioma cells.Results With the results of CCK-8,LN229 showed both dose-dependent and time-dependent manner after γ-irradiation treatment,while in the U251 cells this phenomenon was not obvious.Western blot assay showed that E-cadherin and Claudin were down-regulated after γ-irradiation in both LN229 and U251 cells.However,β-catenin and Vimentin were up-regulated.With the treatment of γ-irradiation(96 h) and curcumin(24 h),in both LN229 and U251 cells,the Hedgehog signaling pathway was suppressed and the EMT process was inhibited either.In combined therapy group,cell invasion and migration capacities were suppressed,as the results of Transwell assay.Conclusion Curcumin can improve the efficiency of γ-irradiation therapy by rescuing the induction of EMT process via the suppression of Hedgehog signaling pathway.
作者
李洋
董佳玮
孙颖
陈辉
孙博
杨东波
LI Yang;DONG Jia-wei;SUN Ying;CHEN Hui;SUN Bo;YANG Dong-bo(Department of Neurosurgery,The Second Affiliated Hospital of Harbin Medical University,Harbin 150081,China)
出处
《哈尔滨医科大学学报》
CAS
2018年第6期539-544,共6页
Journal of Harbin Medical University
基金
哈尔滨医科大学创新科学研究资助项目(2017LCZX46)
关键词
姜黄素
γ-射线
HEDGEHOG信号通路
EMT
curcumin
γ-irradiation
Hedgehog signaling pathway
epithelial-mesenehymal transition
