Glioblastoma stem cells resistant to temozolomide-induced autophagy
Glioblastoma stem cells resistant to temozolomide-induced autophagy
复制标题
胶质母细胞瘤干细胞对替莫唑胺诱导的自噬具有抵抗力
DOI：
10.3760/cma.j.issn.0366-6999.2009.11.004
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发表时间：
2009-06-05
影响因子：
6.1
通讯作者：
Chen Zhong-ping
中科院分区：
文献类型：
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作者：
Fu Jun;Liu Zhi-gang;Chen Zhong-ping
研究背景近年来的研究表明，脑肿瘤中存在一小部分具有原始神经祖细胞特征和肿瘤起始功能的细胞。这些细胞可能是完全根除肿瘤的主要治疗靶点。本研究旨在探讨胶质母细胞瘤干细胞（GSCs）对替莫唑胺（TMZ）的耐药表型及其可能的分子机制。采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑（MTT）法测定TMZ对CD 133（+）和CD 133（-）胶质母细胞瘤细胞的细胞毒作用。通过Western印迹分析自噬相关蛋白（Beclin-1、LC 3和Atg 5）和裂解的caspase-3（p17）。免疫荧光染色检测胶质母细胞瘤细胞中Atg 5、胶质细胞酸性蛋白（GFAP）和CD 133的表达。采用SPSS10.0软件进行统计学分析。对于所有检验，统计学显著性水平设定为P
Background Recent studies have demonstrated the existence of a small fraction of cells with features of primitive neural progenitor cells and tumor-initiating function in brain tumors. These cells might represent primary therapeutic target for complete eradication of the tumors. This study aimed to determine the resistant phenotype of glioblastoma stem cells (GSCs) to temozolomide (TMZ) and to explore the possible molecular mechanisms underlying TMZ resistance.Methods Freshly resected glioblastoma specimen was collected and magnetic isolation of GSCs was carried out using the Miltenyi Biotec CD133 Cell Isolation kit. The cytotoxic effect of TMZ on CD133(+) and CD133(-) glioblastoma cells was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Autophagy-related proteins (Beclin-1, LC3 and Atg5) and cleaved caspase-3 (p17) were analyzed by Western blotting. Immunofluorescent staining was used to detect Atg5, glial fibrillary acidic protein (GFAP) and CD133 expression in glioblastoma cells. Statistical analysis was carried out using SPSS 10.0 software. For all tests, the level of statistical significance was set at P