Glioblastoma is an extremely aggressive glioma, resistant to radio and chemotherapy usually performed with temozolomide. One of the main reasons for glioblastoma resistance to conventional therapies is due to the presence of cancer stem-like cells. These cells could recapitulate some signaling pathways important for embryonic development, such as Sonic hedgehog. Here, we investigated if the inhibitor of the Sonic hedgehog pathway, cyclopamine, could potentiate the temozolomide effect in cancer stem-like cells and glioblastoma cell lines . The viability of glioblastoma cells exposed to cyclopamine and temozolomide treatment was evaluated by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay while the induction of apoptosis was assessed by western blot. The stemness properties of glioma cells were verified by clonogenic and differentiation assay and the expression of stem cell markers were measured by fluorescence microscopy and western blot. The glioblastoma viability was reduced by cyclopamine treatment. Cyclopamine potentiated temozolomide treatment in glioblastoma cell lines by inducing apoptosis through activation of caspase-3 cleaved. Conversely, the combined treatment of cyclopamine and temozolomide potentiated the stemness properties of glioblastoma cells by inducing the expression of SOX-2 and OCT-4. Cyclopamine plays an effect on glioblastoma cell lines but also sensibilize them to temozolomide treatment. Thus, first-line treatment with Sonic hedgehog inhibitor followed by temozolomide could be used as a new therapeutic strategy for glioblastoma patients.

中文翻译：

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环巴明通过音刺猬通路使胶质母细胞瘤细胞对替莫唑胺治疗敏感

胶质母细胞瘤是一种极具侵袭性的胶质瘤，对通常使用替莫唑胺进行的放疗和化疗具有抵抗力。胶质母细胞瘤对传统疗法产生耐药性的主要原因之一是癌症干细胞样细胞的存在。这些细胞可以重现一些对胚胎发育很重要的信号通路，例如 Sonic Hedgehog。在这里，我们研究了音刺猬通路抑制剂环巴明是否可以增强替莫唑胺在癌症干细胞样细胞和胶质母细胞瘤细胞系中的作用。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定评估暴露于环巴明和替莫唑胺处理的胶质母细胞瘤细胞的活力，同时通过蛋白质印迹评估细胞凋亡的诱导。通过克隆形成和分化实验验证神经胶质瘤细胞的干细胞特性，并通过荧光显微镜和蛋白质印迹测量干细胞标记物的表达。环巴明治疗降低了胶质母细胞瘤的活力。 Cyclopamine 通过激活 caspase-3 裂解诱导细胞凋亡，从而增强替莫唑胺对胶质母细胞瘤细胞系的治疗作用。相反，环巴明和替莫唑胺的联合治疗通过诱导 SOX-2 和 OCT-4 的表达增强了胶质母细胞瘤细胞的干细胞特性。环巴明对胶质母细胞瘤细胞系起作用，但也使它们对替莫唑胺治疗敏感。因此，一线治疗采用 Sonic Hedgehog 抑制剂，然后采用替莫唑胺治疗，可作为胶质母细胞瘤患者的新治疗策略。