Wang Zhaotao, Li Yongping, Liu Minyi, Chen Danmin, Lu Jiajie, Ji Yunxiang, Xing Zhou, Wang Yezhong
Institute of Neuroscience, Department of Neurosurgery, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260 China.
J Cancer. 2022 Jan 16;13(4):1203-1213. doi: 10.7150/jca.66674. eCollection 2022.
Glioblastoma (GBM) is a tumor of the central nervous system with an extremely poor prognosis. Stemness and EMT play important roles in GBM progression. 3-benzyl-5-((2-nitrophenoxy) methyl) dihydrofuran-2(3H)-one (3BDO), an autophagy inhibitor, has been reported to exert anti-cancer activities on lung carcinoma. However, the effects of 3BDO on GBM remain unknown. Therefore, the purpose of this study was to explore the effects of 3BDO on GBM and to investigate the underlying molecular mechanisms. CCK-8 experiments and clone formation assays were conducted to determine the level of cell proliferation. Transwell assay was conducted to examine cell migration and invasion abilities. Western blotting and immunofluorescence staining were used to analyze protein expression levels. A xenograft mouse model was used to evaluate the effect of 3BDO . We found that 3BDO inhibited U87 and U251 cell proliferation in a dose-dependent manner. Additionally, 3BDO decreased the degree of sphere formation and levels of stemness markers (sox2, nestin, and CD133) in GSCs. 3BDO also inhibited migration and invasion abilities and suppressed EMT markers (N-cadherin, vimentin, and snail) in GBM cells. Moreover, we found that 3BDO downregulated the expression of survivin in both GBM cells (U87, U251) and GSCs. Furthermore, overexpression of survivin decreased the therapeutic effect of 3BDO on EMT, invasion, migration, and proliferation of GBM cells, as well as decreased the stemness of GSCs. Finally, we demonstrated that 3BDO could inhibit tumor growth in a tumor xenograft mouse model constructed using U87 cells. Similar to the findings, 3BDO decreased the expression of survivin, EMT makers, and the degree of stemness . Our results demonstrate that 3BDO can repress GBM both and via downregulating survivin-mediated stemness and EMT.
胶质母细胞瘤（GBM）是一种中枢神经系统肿瘤，预后极差。干性和上皮-间质转化（EMT）在GBM进展中起重要作用。3-苄基-5-（（2-硝基苯氧基）甲基）二氢呋喃-2（3H）-酮（3BDO）是一种自噬抑制剂，据报道对肺癌具有抗癌活性。然而，3BDO对GBM的影响尚不清楚。因此，本研究的目的是探讨3BDO对GBM的影响，并研究其潜在的分子机制。进行CCK-8实验和克隆形成试验以确定细胞增殖水平。进行Transwell试验以检测细胞迁移和侵袭能力。使用蛋白质免疫印迹和免疫荧光染色分析蛋白质表达水平。使用异种移植小鼠模型评估3BDO的作用。我们发现3BDO以剂量依赖性方式抑制U87和U251细胞增殖。此外，3BDO降低了胶质瘤干细胞（GSCs）中球体形成的程度和干性标志物（sox2、巢蛋白和CD133）的水平。3BDO还抑制GBM细胞的迁移和侵袭能力，并抑制EMT标志物（N-钙黏蛋白、波形蛋白和蜗牛蛋白）。此外，我们发现3BDO下调了GBM细胞（U87、U251）和GSCs中生存素的表达。此外，生存素的过表达降低了3BDO对GBM细胞的EMT、侵袭、迁移和增殖的治疗效果，以及降低了GSCs的干性。最后，我们证明3BDO可以在使用U87细胞构建的肿瘤异种移植小鼠模型中抑制肿瘤生长。与这些发现相似，3BDO降低了生存素、EMT标志物的表达以及干性程度。我们的结果表明，3BDO可以通过下调生存素介导的干性和EMT来抑制GBM。
