Zhu Ting-Zhun, Li Xiao-Ming, Luo Li-Han, Song Zhen-Quan, Gao Xu, Li Zhi-Qing, Su Jing-Yuan, Liang Guo-Biao
Department of Neurosurgery, General Hospital of Shenyang Military Area Command, Shenyang 110840, P.R. China.
Health Care Centre, Shenyang Entry-Exit Inspection and Quarantine Bureau, Shenyang 110016, P.R. China.
Int J Oncol. 2014 Aug;45(2):699-709. doi: 10.3892/ijo.2014.2448. Epub 2014 May 19.
Accumulating evidence indicates that glioblastoma stem-like cells (GSCs) are key factors in tumour development, recurrence and chemoresistance. The impairment of stemness and the enhancement of differentiation contributes to the weakening of radiation and chemotherapy resistance of GSCs. We previously found that β-elemene was an effective anti-glioblastoma agent and chemosensitizer. In this study, we examined the distribution of CD133(+) cells in human glioblastoma tissues by immunohistochemistry. Following treatment with β-elemene, the formation of GSC spheres was investigated by manual counting, the proliferation of GSCs was measured with a Cell Counting Kit-8 (CCK-8) assay, and the dispersion of GSC spheres was observed with an inverted microscope. GSC spheres were treated with β-elemene, and the expression levels of CD133, ATP-binding cassette subfamily G member 2 (ABCG2) and glial fibrillary acidic protein (GFAP) were examined by western blotting. After treatment with β-elemene, the volumes and weights of GSC xenografts were measured, and the expression of CD133, ABCG2 and GFAP was evaluated through immunohistochemistry analysis. After treatment with β-elemene and temozolomide (TMZ), GSC viability was examined by the CCK-8 assay, and the volumes and weights of xenografts were measured. We found that CD133(+) cells were assembled in some vascular walls and also sparsely distributed in other parts of glioblastoma tissues. β-elemene decreased the formation of GSC spheres, dispersed GSC spheres and inhibited the proliferation of GSCs in vitro and in vivo. In the GSC spheres and xenografts treated with β-elemene, the expression of CD133 and ABCG2 was significantly downregulated, and the expression of GFAP increased. Furthermore, the sensitivity of GSCs to TMZ was enhanced in vitro and in vivo. These results suggest that β-elemene impaired the stemness of GSC spheres, promoted their differentiation and sensitized GSCs to TMZ. β-elemene will hopefully become a valuable agent to enhance the effects of radiotherapy and chemotherapy.
越来越多的证据表明，胶质母细胞瘤干细胞（GSCs）是肿瘤发生、复发和化疗耐药的关键因素。干性的损害和分化的增强有助于削弱GSCs的放疗和化疗耐药性。我们之前发现β-榄香烯是一种有效的抗胶质母细胞瘤药物和化学增敏剂。在本研究中，我们通过免疫组化检测了人胶质母细胞瘤组织中CD133(+)细胞的分布。用β-榄香烯处理后，通过手工计数研究GSC球的形成，用细胞计数试剂盒-8（CCK-8）检测GSCs的增殖，并在倒置显微镜下观察GSC球的分散情况。用β-榄香烯处理GSC球，通过蛋白质印迹法检测CD133、ATP结合盒转运体G亚家族成员2（ABCG2）和胶质纤维酸性蛋白（GFAP）的表达水平。用β-榄香烯处理后，测量GSC异种移植瘤的体积和重量，并通过免疫组化分析评估CD133、ABCG2和GFAP的表达。用β-榄香烯和替莫唑胺（TMZ）处理后，通过CCK-8检测GSC的活力，并测量异种移植瘤的体积和重量。我们发现CD133(+)细胞聚集在一些血管壁上，也稀疏分布在胶质母细胞瘤组织的其他部位。β-榄香烯减少了GSC球的形成，分散了GSC球，并在体外和体内抑制了GSCs的增殖。在用β-榄香烯处理的GSC球和异种移植瘤中，CD133和ABCG2的表达明显下调，而GFAP的表达增加。此外，在体外和体内，GSCs对TMZ的敏感性均增强。这些结果表明，β-榄香烯损害了GSC球的干性，促进了它们的分化，并使GSCs对TMZ敏感。β-榄香烯有望成为一种增强放疗和化疗效果的有价值的药物。
