热休克转录因子4（heat shock transcription factor 4,HSF4) 是调控新生儿期晶状体发育的关键转录因子.它参与调控晶状体上皮细胞增生和纤维细胞分化,然而其 作用机理仍不清楚.利用鼠晶状体上皮细胞mLEC/HSF4-/-和mLEC/HA-HSF4b细胞为材料，对HSF4在晶状体上皮细胞中的调控作用进行研究.结果发现，重建的mLEC/HA -HSF4b细胞中，高表达热休克蛋白25（Hsp25）和αB晶体蛋白（αB-crystallin）. 细胞划痕实验和Transwell 细胞迁移实验结果表明，mLEC/HSF4-/-细胞明显比 mLEC/HA-HSF4b细胞的迁徙速度慢（t检验, P=0.0001).应用免疫印迹和半定量RT- PCR实验发现，HSF4b能上调 RhoA, Rac1 和 CDC42的表达.但后者的表达不受Hsp25 和αB-crystallin调控. 由此推论，HSF4b通过调控RhoA, Rac1 和CDC42 的表达参与调控晶状体上皮细胞向后房的迁徙. 这一新发现对解释HSF4b调控晶状体发育提供了有力的证据.
Abstract
Heat shock transcription factor 4 (HSF4) is a key regulator of postnatal lens development by involving the regulation of lens epithelial cell proliferation and fibroblast differentiation. We found that the expression of Hsp25 and αB-crystallin was up-regulated in HSF4b- reconstituted mLEC/HA-HSF4b cells. The results of wound-healing and Transwell assays indicated that the migration of mLEC/HA-HSF4b cells was significantly faster than that of mLEC/HSF4-/-cells (P=0.0001). The immunoblotting and semi-quantity RT-PCR data showed that HSF4b up- regulated the expression of RhoA, Rac1 and CDC42 and correlated with the Hsp25 and αB-crystallin levels,except for CDC42. We hypothesized that HSF4b might regulate the migration of anterior epithelial cell to posterior by increasing the expression of RhoA and Rac1.
关键词
热休克转录因子4 /
晶状体上皮细胞 /
RhoA /
Rac1 /
热休克蛋白25 /
αB-晶体蛋白
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Key words
123 /
αB-crystallin
heat shock factor 4 ( HSF4 ) /
mouse lens epithelial cell /
RhoA /
Rac1 /
heat shock protein 25 (Hsp25 ) /
αB-crystallin
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中图分类号：
Q78
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参考文献
[1] Bu 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
[2] Min J N, 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
[3] Mitchell D C, Bryan B A, Liu J P, et al. Developmental expression of three small GTPases in the mouse eye[J]. Mol Vis, 2007, 13(13):1144-1153
[4] Zhang J, Hu Y Z, Xueli L, et al. The inhibition of CMV promoter by heat shock factor 4b is regulated by Daxx [J]. Int J Biochem Cell Biol, 2010，42(10) :1698-1707
[5]Hu Y Z, Zhang J, Li S, et al. The transcription activity of heat shock factor 4b is regulated by FGF2[J]. Int J Biochem Cell Biol, 2012, 45(2):317-325
[6] Ridley A J. Rho GTPases and cell migration [J]. J Cell Sci, 2001, 114(15):2713-2722
[7]Hu Y, Mivechi N F. Association and regulation of heat shock transcription factor 4b with both extracellular signal-regulated kinase mitogen-activated protein kinase and dual-specificity tyrosine phosphatase DUSP26[J]. Mol Cell Biol, 2006, 26(8): 3282-3294
[8]Shi X, Cui B, Wang Z, et al. Removal of Hsf4 leads to cataract development in mice through down- regulation of gamma S-CRYSTALLIN and Bfsp expression[J]. BMC Mol Biol, 2009, 19 (10): 10
[9] Mou L, Xu J Y, Li W , et al. Identification of vimentin as a novel target of HSF4 in lens development and cataract by proteomic analysis[J]. Invest Ophthalmol Vis Sci, 2010, 51(1): 396-404
[10] Maddala R, Reddy V N, Epstein D L ,et al. Growth factor induced activation of Rho and Rac GTPases and actin cytoskeletal reorganization in human lens epithelial cells[J]. 2003 , 9:329-336
[11]谭运年,成海恩,金亚平. Rac1、Rac2参与调节人单核细胞趋化和NADPH氧化酶活性[J].中国生物化学与分子生物学报(Tan YN, Cheng HN, Jin YP. Rac1 and Rac2 are involved in chemotaxis and NADPH oxidase activity in human monocytes[J]. Chin J Biochem Mol Biol) , 2011，27( 9) : 858-863
[12] Plageman T F Jr, Chauhan B K, Yang C , et al. A Trio-RhoA-Shroom3 pathway is required for apical constriction and epithelial invagination[J]. 2011, 138(23):5177-5188
[13]Chauhan B K, Lou M, Zheng Y, et al. Balanced Rac1 and RhoA activities regulate cell shape and drive invagination morphogenesis in epithelia [J]. Proc Natl Acad Sci U S A, 2011, 108(45): 18289-18294
[14] Maddala R, Chauhan BK, Walker C , et al. Rac1 GTPase-deficient mouse lens exhibits defects in shape, suture formation, fiber cell migration and survival[J]. Dev Biol, 2011, 360(1):30-43
[15] Chauhan B K, Disanza A, Choi S Y, et al. Cdc42- and IRSp53-dependent contractile filopodia tether presumptive lens and retina to coordinate epithelial invagination [J].Development,2009,136(21):3657- 3667
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脚注
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基金
国家自然科学基金资助(No. 30871299 ,No.81270985)
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