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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。
Objective To study the role of microtubule cross-connection factor 1 (MACF1) in response to TMZ stimulation in glioblastoma. Methods After stimulating the human glioblastoma cell line U87 cells with temozolomide, the protein expression and intracellular localization were detected by Western blot, RT-PCR and immunofluorescence. Interference of MACF1 expression by RNA interference techniques. The expression of MACF1 in glioblastoma after TMZ stimulation in vivo was detected by subcutaneous tumor formation and immunohistochemistry in nude mice. Results The expression of MACF1 in glioblastoma cells was up-regulated after temozolomide stimulation (about 2-fold difference) and intracellular distribution (P <0.01) both in vitro and in vivo. Glioblastoma cells knocked out of MACF1 expression reduced their proliferative capacity by about 45% (P <0.01) after TMZ stimulation. Temozolomide induced glioblastoma MACF1 expression increased at the same time, the cytoskeleton occurred recombinant. Conclusion MACF1 may be a potential target for the treatment of glioblastoma.