HOXD13调控SOX9基因可能参与先天足部软组织畸形的发生

doi:10.13418/j.issn.1001-165x.2020.06.013

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (6) : 691-696. DOI: 10.13418/j.issn.1001-165x.2020.06.013

实验研究

HOXD13调控SOX9基因可能参与先天足部软组织畸形的发生

王正东¹，　颜南¹，　刘丽英²，　李舒¹，　陈尧¹，　孔锶慧¹，　李培源¹，　金春莲³

作者信息 +

Regulation of SOX9 by HOXD13 may be related to the occurrence of congenital foot malformation

WANG Zheng-dong¹, YAN Nan¹, LIU Li-ying², LI Shu¹, CHEN Yao¹, KONG Si-hui¹,LI Pei-yuan¹, JIN Chun-lian³

Author information +

文章历史 +

摘要

目的　研究HOXD13与SOX9基因在单纯性马蹄内翻足（idiopathic congenital talipes equinovarus，ICTEV）足部软组织畸形发生中的分子机制。方法　软件预测SOX9基因5′上游3个HOXD13结合位点，构建SOX9基因上游不同长度片段以及结合位点野生和突变序列载体，并测定其荧光素酶活性。在体内用ChIP验证HOXD13与SOX9基因启动子区3位点结合作用。干扰HOXD13基因表达，qRT-PCR检测HOXD13和SOX9表达水平的改变。结果　荧光素酶检测发现在SOX9基因5′上游-1158至-770 bp（位点1）和-512至-368 bp之间（位点3）为负调控区；-770至-512 bp之间（位点2）是正调控区。ChIP检测显示HOXD13蛋白可与SOX9基因位点3结合。干扰HOXD13表达，SOX9基因mRNA表达上调。结论　HOXD13结合SOX9基因5′上游负调控区可上调SOX9基因表达。干扰HOXD13会上调SOX9基因表达，高表达SOX9基因可能与足踝部软组织的挛缩发生相关。

Abstract

Objective　To study the molecular mechanism of HOXD13 and SOX9 genes in the occurrence of foot soft tissue malformation of idiopathic congenital talipes equinovarus (ICTEV).　Methods 3 HOXD13 binding sites upstream of 5 'of SOX9 gene were predicted by the software, and luciferase activity of serial truncated and mutant vector in promoter region of SOX9 gene was detected. The binding effect of HOXD13 to the 3 sites of SOX9 gene promoter was verified by ChIP in vivo. The expression levels of HOXD13 and SOX9 were detected by qRT-PCR.　Results　A negative regulatory region between -1158 to -770 bp (site 1) and -512 to -368 bp (site 3) in the upstream of SOX9 gene 5' was found by the Luciferase. Between -770 and -512bp (locus 2) was a positive regulatory region. ChIP results showed that HOXD13 could bind to SOX9 locus 3.The mRNA expression of SOX9 gene was up-regulated by interference of HOXD13 expression.　Conclusions　The expression of SOX9 gene was up-regulated by HOXD13 in combination with the negative regulation region of SOX9 gene 5' upstream. The interference of HOXD13 up-regulated SOX9 gene expression may be related to the occurrence of soft tissue contracture on the ankle of foot.

导出引用

王正东, 颜南, 刘丽英, 李舒, 陈尧, 孔锶慧, 李培源, 金春莲. HOXD13调控SOX9基因可能参与先天足部软组织畸形的发生[J]. 中国临床解剖学杂志. 2020, 38(6): 691-696 https://doi.org/10.13418/j.issn.1001-165x.2020.06.013

WANG Zheng-dong, YAN Nan, LIU Li-ying, LI Shu, CHEN Yao, KONG Si-hui, LI Pei-yuan, JIN Chun-lian. Regulation of SOX9 by HOXD13 may be related to the occurrence of congenital foot malformation[J]. Chinese Journal of Clinical Anatomy. 2020, 38(6): 691-696 https://doi.org/10.13418/j.issn.1001-165x.2020.06.013

中图分类号： Q291

参考文献

[1] Ponseti IV, Campos J. The classic: observations on pathogenesis and treatment of congenital clubfoot. 1972[J]. Clin Orthop Relat Res, 2009, 467(5): 1124-1132.
[2] Wang ZD, Yan N, Liu LY, et al. SOX9 overexpression plays a potential role in idiopathic congenital talipes equinovarus[J]. Mol Med Rep, 2013, 7(3): 821-825.
[3] 王正东, 颜南, 王效杰, 等. Sox9基因在马蹄内翻足大鼠模型足踝软骨组织表达的实验研究 [J]. 沈阳医学院学报, 2012, 14(4): 199-201.
[4] Zhang P, Jimenez SA, Stokes DG. Regulation of human COL9A1 gene expression. Activation of the proximal promoter region by SOX9[J]. J Biol Chem, 2003, 278(1): 117-123.
[5] 赵宁, 金春莲, 刘丽英, 等. COL1A1基因转录调控序列变异与单纯性马蹄内翻足的相关性 [J]. 遗传, 2008, 30(6): 723-727.
[6] Ippolito E, De Maio F, Mancini F, et al. Leg muscle atrophy in idiopathic congenital clubfoot: is it primitive or acquired[J]? J Child Orthop, 2009, 3(3): 171-178.
[7] Liu LY, Jin CL, Jiang L, et al. [Expression of COL9A1 gene and its polymorphism in children with idiopathic congenital talipes equinovarus][J]. Zhongguo Dang Dai Er Ke Za Zhi, 2011, 13(6): 478-481.
[8] Wang LL, Fu WN, Li ZG, et al. Research of HOXD13 and FHL1 in idiopathic congenital talipes equinovarus[J]. Yi Chuan, 2008, 30(1): 46-50.
[9] Zaib T, Ji W, Saleem K, et al. A heterozygous duplication variant of the HOXD13 gene caused synpolydactyly type 1 with variable expressivity in a Chinese family[J]. BMC Med Genet, 2019, 20(1): 203.
[10] Wang LL, Jin CL, Liu LY, et al. [Analysis of association between 5' HOXD gene and idiopathic congenital talipes equinovarus][J]. Zhonghua Yi Xue Yi Chuan Xue Za Zhi, 2005, 22(6): 653-656.
[11] Ibrahim DM, Tayebi N, Knaus A, et al. A homozygous HOXD13 missense mutation causes a severe form of synpolydactyly with metacarpal to carpal transformation[J]. Am J Med Genet A, 2016, 170(3): 615-621.
[12]Salsi V, Vigano MA, Cocchiarella F, et al. Hoxd13 binds in vivo and regulates the expression of genes acting in key pathways for early limb and skeletal patterning[J]. Dev Biol, 2008, 317(2): 497-507.
[13]Wang LL, Fu WN, Li-Ling J, et al. HOXD13 may play a role in idiopathic congenital clubfoot by regulating the expression of FHL1[J]. Cytogenet Genome Res, 2008, 121(3-4): 189-195.
[14]Cao DH, Jin CL, Ren MH, et al. The expression of Gli3, regulated by HOXD13, may play a role in idiopathic congenital talipes equinovarus[J]. BMC Musculoskelet Disord, 2009, 10: 142.