Objective: To investigate the role of microtubule-actin crosslinking factor 1 (MACF1) in the response of glioma cells to temozolomide (TMZ).
Methods: TMZ was applied to a human gliomablastoma cell line (U87) and changes in the protein expression and cellular localization were determined with Western blot, RT-PCR, and immunofluorescence. The responses of the cells with MACF1 expression knockdown by RNA interference to TMZ were assessed. TMZ-induced effects on MACF1 expression were also assessed by immunohistochemistry in a nude mouse model bearing human glioblastoma xenografts.
Results: TMZ resulted in significantly increased MACF1 expression (by about 2 folds) and changes in its localization in the gliomablastoma cells both in vitro and in vivo (P<0.01). Knockdown of MACF1 reduced the proliferation (by 45%) of human glioma cell lines treated with TMZ (P<0.01). TMZ-induced changes in MACF1 expression was accompanied by cytoskeletal rearrangement.
Conclusion: MACF1 may be a potential therapeutic target for glioblastoma.
目的: 研究微管微丝交连因子1（MACF1）在胶质母细胞瘤应对TMZ刺激中的作用。
方法: 替莫唑胺刺激人类胶质母细胞瘤细胞系U87细胞后，通过Western blot、RT-PCR和免疫荧光检测其蛋白表达和细胞内定位。通过RNA干扰技术干扰MACF1的表达。通过裸鼠皮下成瘤和免疫组化检测在活体中TMZ刺激后胶质母细胞瘤中MACF1的表达。
结果: 在体外培养和体内成瘤中均发现替莫唑胺刺激后胶质母细胞瘤MACF1的表达上调（差异约2倍），胞内分布改变（P < 0.01）。敲除MACF1表达的胶质母细胞瘤细胞在TMZ刺激后其增殖能力下调约45%（P < 0.01）。替莫唑胺诱导胶质母细胞瘤MACF1表达上调的同时，其细胞骨架发生重组。
结论: MACF1可能成为胶质母细胞瘤治疗的一个潜在靶点。