Hsf4-/-小鼠晶状体永生化上皮细胞系建立的研究  

Establishment of mouse lens epithelial cell lines with the genotype of Hsf4-/-

作　　者：张军[1] 马增翼[2] 王悦玲[1] 王继燕[1] 胡延忠[1] 

机构地区：[1]河南大学医学院细胞与分子免疫学重点实验室,开封475000 [2]复旦大学附属华山医院神经外科

出　　处：《中华眼科杂志》2013年第11期1029-1031,共3页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目（30871299）

摘　　要：目的 建立Hsf4-/-小鼠的晶状体永生化上皮细胞系.方法 实验研究.取新生P6天的Hsf4-/-小鼠晶状体原代培养晶状体上皮细胞（MLEC）,用SV40,T-抗原转染,C418筛选建立永生化细胞系MLEC/Hsf4-/-,免疫印迹法检测α-A晶状体蛋白表达.结果 倒置相差显微镜下观察可见细胞贴壁生长,有伪足伸出,多角形,大小均匀,细胞膜边界清晰,细胞质透亮.消化后的细胞呈圆形透亮,胞质丰富.免疫印迹法检测MLEC/Hsf4-/-细胞（两个克隆）中的α-A晶状体蛋白（α-A-crystallin）有表达.转染Hsf4b蛋白后Hsp25蛋白表达上调.结论 采用组织块接种法获得小鼠晶状体上皮细胞后,转染SV40大T-抗原,G418筛选可成功建立永生化细胞系MLEC/Hsf4-/-.Objective To establish mouse lens epithelial cell lines with the genotype of Hsf4-/-.Methods The expended mouse lens epithelial cells,which were generated from P6 Hsf4-deficient mouse lens epithelia,were immortalized with SV40-T-antigen and named MLEC/Hsf4-/-cell.The expression of alpha A-crystallin was immunoblotted.Hsf4b cDNA was reconstituted by transiently transfection.Results The SV40-immortalized cells were in adherent growth mode with spindle morphology,pseudopodia,clear nuclear bondary membrane and cytoplasm translucent.Immunoblotting results indicated that the lens biomarker protein alpha A-cryatllin was expressed in MLEC/Hsf4-/-cells.Reconstitution of Hsf4b into MLEC/Hsf4-/-cells upregulated the expression of Hsp25.Conclusions The SV40-immortalized MLEC/ Hsf4-/-cells have the lens epithelial characteristics and could be used as a tool for studying the signal transduction in vitro.

关 键 词：DNA结合蛋白质类 转录因子 晶体 上皮细胞 细胞系 免疫印迹法 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]