人胶质瘤细胞的激素诱导型细胞凋亡  

Hormone-Induced Apoptosis of Human Glioma Cells

作　　者：孙蕾[1] 王会信[1] 郑莉[1] 王芳[1] 刘农乐[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《中国肿瘤生物治疗杂志》1999年第1期35-38,共4页Chinese Journal of Cancer Biotherapy

摘　　要：目的:利用β-雌二醇受体的性质,构建Fas与β-雌二醇受体融合基因,使转染该融合基因的胶质瘤细胞能在激素的诱导下发生凋亡.方法:利用PCR技术及基因重组手段,将人Fas基因的跨膜区、胞内区和人β-雌二醇受体的激素结合结构域基因片断融合,并插入高效真核表达载体peDNA3.脂质体法转染人胶质瘤细胞BT325.结果:加入G4186周后筛选出的转化细胞,经Westem blot检测证明,该融合基因在BT325细胞中得到较高表达.MTT法检测表明,在培养基中加入一定浓度的β-雌二醇.可有效地杀死转化细胞,IC_(50)为10^(-9)mol/L DNA Ladder检测进一步证明,转化细胞被诱导发生凋亡.结论:Fas与β-雌二醇受体融合基因转染的胶质瘤细胞,其调亡严格受激素调控.Objective: Using the property of (β-estrogen receptor, Fas/β-estrogen receptor fusion gene was constructed which can lead to hormone-induced apoptosis after transfected into glioma cells. Methods: The transmembrane domain, cytoplasmic domain of human Fas gene was fused with the HBD gene fragment of human (β-estrogen receptor by PCR and gene recombination techniques, and was then inserted into eukaryotic vector pcDNA3. Human glioma cells BT325 were transfected with the recombinant plasmid by lipofectamine-rnediated gene transfection. Results: After selection with C418 (or six weeks, transformants expressing the fusion gene were selected out and identified by Western blot. MTT detection showed that (β-estradiol had cytoxic effect on the transformants with IC_(50) of about 10^(-9) mol/L. DNA Ladder detection showed that the transformants could be effectively induced to apoptosis. Conclusion: Fas/β-estrogen receptor fusion gene transfected glioma cells can be induced to apoptosis in a tight estrogen indepent manner.

关 键 词：细胞凋亡 胶质瘤细胞 激素诱导 

分 类 号：R730.264[医药卫生—肿瘤]