期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

Hsf4-/-小鼠晶状体永生化上皮细胞系建立的研究 被引量：1

Establishment of mouse lens epithelial cell lines with the genotype of Hsf4-/-
原文传递
导出
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.
作者 张军 马增翼 王悦玲 王继燕 胡延忠
机构地区 河南大学医学院细胞与分子免疫学重点实验室 复旦大学附属华山医院神经外科
出处 《中华眼科杂志》 CAS CSCD 北大核心 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）
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 DNA-binding proteins Transcription factors Lens, crystalline Epithelial cells Cell line Immunoblotting
分类号 R329 [医药卫生—人体解剖和组织胚胎学]
  • 相关文献

参考文献2

二级参考文献22

  • 1Nakai A. New aspects in the vertebrate heat shock factor system: Hsf3 and Hsf4[J]. Cell Stress Chaperones, 1999, 4(2) : 86-93.
  • 2Cotto J J , Kline M, Morimoto RI. Activation of heat shock factor 1 DNA binding precedes stress-induced serine phosphorylation. Evidence for a muhistep pathway of regulation [ J ]. J Biol Chem, 1996, 271 (7) : 3355 -3358.
  • 3Fujimoto M, Izu H, Seki K, et al. HSF4 is required for normal cell growth and differentiation during mouse lens development[ J]. EMBO J, 2004, 23(21): 4297-4306.
  • 4Smaoui N, Behaief O, Benhamed S, et al. A homozygous splice mutation in the HSF4 gene is associated with an autosomal recessive congenital cataract [ J ]. Invest Ophthalmol Vis Sci, 2004, 45 ( 8 ) : 2716 - 2721.
  • 5Min JN, Zhang Y, Moskophidis D, et al. Unique contribution of heat shock transcription factor 4 in ocular lens development and fiber cell differentiation[J]. Genesis, 2004, 40(4): 205-217.
  • 6Tanabe M, Sasai N, Nagata K, et al. The mammalian HSF4 gene generates both an activator and a repressor of heat shock genes by alternative splicing [ J ]. J Biol Chem, 1999, 274 ( 39 ) : 27545 - 27856.
  • 7Hu Y, Mivechi NF. Association and regulation of heat shock transcription factor 4b with both extracellular sigual-regulated kinase mitogenactivated protein kinase and dual-specificity tyrosine phosphatase DUSP26[J]. Mol Cell Biol, 2006, 26(8) : 3282 -3294.
  • 8Hietakangas V, Anckar J, Blomster HA, et al. PDSM, a motif for phosphorylation-dependent SUMO modification [ J]. Proc Natl Acad Sci USA, 2006, 103(1) : 45 -50.
  • 9Frejtag W, Zhang Y, Dai R, et al. Heat shock factor-4 (HSF-4a) represses basal transcription through interaction with TFIIF[ J]. J Biol Chem, 2001, 276(18): 14685-14694.
  • 10Tu N, Hu Y, Mivechi NF. Heat shock transcription factor (Hsf) -4b recruits Brgl during the G1 phase of the cell cycle and regulates the expression of heat shock proteins[J]. J Cell Biochem, 2006, 98 (6) : 1528 -1542.

共引文献9

同被引文献17

  • 1吴明星,吴开力,卞庆宁,姬红培,王忠浩,刘奕志.年龄相关性白内障晶体上皮细胞的基因表达谱变化的初步分析[J].中国病理生理杂志,2006,22(7):1429-1434. 被引量：4
  • 2Min JN, Zhang Y, Moskophidis D, et al. Unique contri- bution of heat shock transcription factor 4 in ocular lens development and fiber cell differentiation [ J ]. Genesis, 2004, 40(4) :205-217.
  • 3Merath K, Ronchetti A, Sidjanin DJ. Functional analysis of HSF4 mutations found in patients with autosomal reces- sive congenital cataracts [ J ]. Invest Ophthalmol Visual Sei, 2013, 54(10):6646-6654.
  • 4Zhang J, Ma Z, Wang J, et al. Regulation of Hsf4b nu- clear translocation and transcription activity by phosphoryl- ation at threonine 472[ J]. Bioehim Biophys Aeta, 2014, 1843(3) :580-589.
  • 5Hu Y, Zhang J, Wang C, et al. The transcription activity of heat shock factor 4b is regulated by FGF2 [ J ]. Int J Biochem Cell Biol, 2013, 45(2) :317-325.
  • 6Nakai A, Tanabe M, Kawazoe Y, et al. HSF4, a new member of the human heat shock factor family which lacks properties of a transcriptional activator[ J]. Mol Cell Biol, 1997, 17(1) :469-481.
  • 7Ke T, Wang QK, Ji B, et al. Novel HSF4 mutation cau- ses congenital total white cataract in a Chinese family[ J]. Am J Ophthalmol, 2006, 142(2) :298-303.
  • 8Bu L, Jin Y, Shi Y, et al. Mutant DNA-binding domain of HSF4 is associated with autosomal dominant lamellar and Marner cataract[J]. Nat Genet, 2002, 31 (3) :276- 278.
  • 9Cui X, Zhang J, Du R, et al. HSF4 is involved in DNA damage repair through regulation of Rad51 [ J ]. Biochim Biophys Acta, 2012, 8(15) : 1308-1315.
  • 10Enoki Y, Mukoda Y, Furutani C, et al. DNA-binding and transcriptional activities of human HSF4 containing muta- tions that associate with congenital and age-related cata- racts[ J ]. Biochim Biophys Acta, 2010, 1802 (9) :749- 753.

引证文献1

中华眼科杂志
2013年 第11期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部