Zhao Wenyang, Peng Fei, Shu Mengting, Liu Huailei, Hou Xu, Wang Xiaoxiong, Ye Junyi, Zhao Boxian, Wang Kaikai, Zhong Chen, Xue Linmeng, Gao Ming, Liu Yaohua, Zhao Shiguang
Department of Neurosurgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Institute of Brain Science, Harbin Medical University, Harbin, China.
Cell Physiol Biochem. 2017;44(4):1381-1395. doi: 10.1159/000485535. Epub 2017 Nov 30.
BACKGROUND/AIMS: Glioma is the most devastating cancer in the brain and has a poor prognosis in adults. Therefore, there is a critical need for novel therapeutic strategies for the management of glioma patients. Isogambogenic acid, an active compound extracted from the Chinese herb Garcinia hanburyi, induces autophagic cell death.
Cell viability was detected with MTT assays. Cell proliferation was assessed using the colony formation assay. Morphological changes associated with autophagy and apoptosis were tested by TEM and Hoechst staining, respectively. The apoptosis rate was measured by flow cytometry. Western blot, immunofluorescence and immunohistochemical analyses were used to detect protein expression. U87-derived xenografts were established for the examination of the effect of isogambogenic acid on glioma growth in vivo.
Isogambogenic acid induced autophagic death in U87 and U251 cells, and blocking late-stage autophagy markedly enhanced the antiproliferative activities of isogambogenic acid. Moreover, we observed the activation of AMPK-mTOR signalling in isogambogenic acid-treated glioma cells. Furthermore, the activation of AMPK or the inhibition of mTOR augmented isogambogenic acid-induced autophagy. Inhibition of autophagy attenuated apoptosis in isogambogenic acid-treated glioma cells. Finally, isogambogenic acid inhibited the growth of U87 glioma in vivo.
Isogambogenic acid inhibits the growth of glioma via activation of the AMPK-mTOR signalling pathway, which may provide evidence for future clinical applications in glioma therapy.
背景/目的：胶质瘤是最具侵袭性的脑癌，成人患者预后较差。因此，迫切需要新的治疗策略来管理胶质瘤患者。异藤黄酸是从中药藤黄中提取的一种活性化合物，可诱导自噬性细胞死亡。
采用MTT法检测细胞活力。使用集落形成试验评估细胞增殖。分别通过透射电镜（TEM）和Hoechst染色检测与自噬和凋亡相关的形态学变化。通过流式细胞术测量凋亡率。采用蛋白质免疫印迹法、免疫荧光法和免疫组织化学分析法检测蛋白质表达。建立U87细胞来源的异种移植模型，以检测异藤黄酸对体内胶质瘤生长的影响。
异藤黄酸诱导U87和U251细胞发生自噬性死亡，阻断晚期自噬显著增强了异藤黄酸的抗增殖活性。此外，我们观察到异藤黄酸处理的胶质瘤细胞中AMPK-mTOR信号通路被激活。此外，激活AMPK或抑制mTOR可增强异藤黄酸诱导的自噬。抑制自噬可减弱异藤黄酸处理的胶质瘤细胞的凋亡。最后，异藤黄酸在体内抑制U87胶质瘤的生长。
异藤黄酸通过激活AMPK-mTOR信号通路抑制胶质瘤生长，这可能为其未来在胶质瘤治疗中的临床应用提供依据。
