摘要
目的 在人胶质母细胞瘤细胞系SHG 44细胞中检测到芳香化酶细胞色素P45 0 (AROM )表达的基础上探讨AROM对胶质瘤细胞生长增殖及分化的作用。方法 通过构建反义AROM真核表达载体 ,转染SHG 44细胞 ,利用流式细胞仪、MTT法、免疫荧光染色及激光共聚焦扫描显微镜 (CLSM)等技术观察封闭AROM基因表达后细胞生长曲线、细胞周期及中间丝蛋白GFAP和Vi mentin表达的变化。结果 成功构建了反义AROM真核表达载体PCI/as AROM并使AROM的表达封闭达 5 0 %以上。SHG 44细胞在转染了PCI/as AROM和空载体PCI neo后 ,生长曲线、细胞周期无明显变化 ,但GFAP免疫荧光强度减弱而Vimentin免疫荧光强度增强。结论 反义AROM可能对SHG 44细胞增殖无明显影响但对神经胶质瘤细胞的分化有一定的抑制作用。
Objective To investigate the effect of cytochrome P450 aromatase gene on the growth,proliferation and differentiation of malignant glioma cell line SHG 44. Methods Gene recombination technique and gene transfection were used to block the expression of aromatase gene. Changes of cell growth curve, cell cycle and expressions of glial fibrillary acid protein (GFAP) and Vimentin were detected by flow cytometry, MTT method, immunofluorescent staining and confocal laser scanning microscopy (CLSM) . Results Antisense aromatase mammalian expression vector PCI/as AROM was constructed in success. The expression of aromatase was blocked in the cells (>50%) by gene transfection. No obvious difference was found between PCI/as AROM and PCI neo tranfected cells in growth velocity and proliferation cycle. The intensity of Vimentin immunofluorecence was increased, while the intensity of GFAP immunofluorescence was decreased. Conclusion Results suggest that antisense aromatase may be involved in inhibiting the differentiation of cell line SHG 44 but have no effect on cell proliferation.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2002年第8期928-930,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 39870 2 74 )
重庆市科委应用基础研究资助项目 ( 2 0 0 1)
