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

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

1.沈阳医学院，沈阳 110034；　2.沈阳市妇婴医院，沈阳 110011；　3.中国医科大学，沈阳 110001

收稿日期:2020-03-04 出版日期:2020-11-25 发布日期:2020-12-08
作者简介:王正东（1977-），男，辽宁辽中人，博士，副教授，硕士生导师，主要从事神经肌肉疾病分子机制的研究，E-mail：wzd_2008@163.com
基金资助:
辽宁省科学技术计划项目(2017225060)

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

1. Shenyang Medical College，Shenyang 11034; 2. Shenyang Maternal and Infant Hospital, Shenyang 110011; 3. China Medical University, Shenyang 110001, China

Received:2020-03-04 Online:2020-11-25 Published:2020-12-08

摘要/Abstract

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。

关键词: 先天足部畸形, 　HOXD13, 　SOX9, 　发育, 　调控

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.

Key words: Congenital foot deformity, 　HOXD13, 　SOX9, 　Development, 　Regulation

中图分类号:

Q291

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.

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.

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HOXD13调控SOX9基因可能参与先天足部软组织畸形的发生

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

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