EZH2对先天性巨结肠结肠组织中GFRα1表达的影响

赵凡, 周崇高, 许光, 马体栋, 夏仁鹏, 李碧香

中国当代儿科杂志 ›› 2019, Vol. 21 ›› Issue (10) : 1033-1037.

PDF(1462 KB)
HTML
PDF(1462 KB)
HTML
中国当代儿科杂志 ›› 2019, Vol. 21 ›› Issue (10) : 1033-1037. DOI: 10.7499/j.issn.1008-8830.2019.10.015
论著·临床研究

EZH2对先天性巨结肠结肠组织中GFRα1表达的影响

  • 赵凡, 周崇高, 许光, 马体栋, 夏仁鹏, 李碧香
作者信息 +

Effect of enhancer of zeste homolog 2 on the expression of glial cell line-derived neurotrophic factor family receptor α-1 in the colon tissue of children with Hirschsprung's disease

  • ZHAO Fan, ZHOU Chong-Gao, XU Guang, MA Ti-Dong, XIA Ren-Peng, LI Bi-Xiang
Author information +
文章历史 +

摘要

目的 通过检测先天性巨结肠(HSCR)患儿结肠组织内胶质细胞源性神经营养因子受体α1(GFRα1)及果蝇Zeste基因增强子人类同源物2(EZH2)的表达水平,探讨EZH2在调控GFRα1基因表达及HSCR发病过程中的作用。方法 随机选取24例行巨结肠根治术的HSCR患儿,取痉挛段结肠组织为试验组。以同期18例因新生儿坏死性小肠结肠炎而接受手术治疗的患儿,取手术切除的坏死结肠组织作为对照组。利用实时荧光定量PCR及Western blot检测两组结肠组织内GFRα1、EZH2的表达水平。将人神经母细胞瘤细胞SHSY5Y分为EZH2过表达组和阴性对照组,EZH2过表达组转染pCMV6-EZH2质粒,阴性对照组转染pCMV6质粒,检测转染后细胞中GFRα1、EZH2的表达水平。结果 与对照组相比,试验组GFRα1及EZH2 mRNA和蛋白的表达水平降低(P < 0.05),且EZH2蛋白表达水平与GFRα1蛋白呈正相关(r=0.606,P=0.002)。与阴性对照组相比,EZH2过表达组EZH2及GFRα1表达水平明显上调(P < 0.05)。结论 HSCR患儿结肠组织中EZH2低表达可能是GFRα1表达不足,诱发HSCR的原因之一。

Abstract

Objective To study the expression levels of glial cell line-derived neurotrophic factor family receptor α-1 (GFRα1) and enhancer of zeste homolog 2 (EZH2) in the intestinal tissue of children with Hirschsprung's disease (HSCR), as well as the role of EZH2 in the regulation of GFRα1 gene expression and the pathogenesis of HSCR. Methods The samples of colon tissue with spasm from 24 children with HSCR after radical treatment of HSCR were selected as the experimental group, and the samples of necrotized colon tissue from 18 children with neonatal necrotizing enterocolitis after surgical resection were selected as the control group. Real-time PCR and Western blot were used to measure the expression levels of GFRα1 and EZH2 in colon tissue in both groups. Human neuroblastoma SH-SY5Y cells were divided into an EZH2 over-expression group and a negative control group. The cells in the EZH2 over-expression group were transfected with pCMV6-EZH2 plasmid, and those in the negative control group were transfected with pCMV6 plasmid. The expression levels of EZH2 and GFRα1 were measured after transfection. Results Compared with the control group, the experimental group had significant reductions in the mRNA and protein expression levels of GFRα1 and EZH2 in colon tissue (P < 0.05), and the protein expression of EZH2 was positively correlated with that of GFRα1 (r=0.606, P=0.002). Compared with the negative control group, the EZH2 over-expression group had significant increases in the expression levels of EZH2 and GFRα1 after SH-SY5Y cells were transfected with EZH2 over-expression plasmid (P < 0.05). Conclusions Low expression of EZH2 in the colon tissue of children with HSCR may be one of the causes of inadequate expression of GFRα1 and onset of HSCR.

关键词

先天性巨结肠 / 胶质细胞源性神经营养因子受体α1 / 果蝇Zeste基因增强子人类同源物2 / 儿童

Key words

Hirschsprung's disease / Glial cell line-derived neurotrophic factor family receptor α1 / Enhancer of zeste homolog 2 / Child

引用本文

导出引用
赵凡, 周崇高, 许光, 马体栋, 夏仁鹏, 李碧香. EZH2对先天性巨结肠结肠组织中GFRα1表达的影响[J]. 中国当代儿科杂志. 2019, 21(10): 1033-1037 https://doi.org/10.7499/j.issn.1008-8830.2019.10.015
ZHAO Fan, ZHOU Chong-Gao, XU Guang, MA Ti-Dong, XIA Ren-Peng, LI Bi-Xiang. Effect of enhancer of zeste homolog 2 on the expression of glial cell line-derived neurotrophic factor family receptor α-1 in the colon tissue of children with Hirschsprung's disease[J]. Chinese Journal of Contemporary Pediatrics. 2019, 21(10): 1033-1037 https://doi.org/10.7499/j.issn.1008-8830.2019.10.015

参考文献

[1] Amiel J, Sproat-Emison E, Garcia-Barcelo M, et al. Hirschsprung disease, associated syndromes and genetics:a review[J]. J Med Genet, 2008, 45(1):1-14.
[2] Alves MM, Sribudiani Y, Brouwer RW, et al. Contribution of rare and common variants determine complex diseases-Hirschsprung disease as a model[J]. Dev Biol, 2013, 382(1):320-329.
[3] 甄亚琴,徐纪荣.先天性巨结肠的研究进展[J].中国当代医药, 2015, 22(34):21-24.
[4] Lui VC, Samy ET, Sham MH, et al. Glial cell line-derived neurotrophic factor family receptors are abnormally expressed in aganglionic bowel of a subpopulation of patients with Hirschsprung's disease[J]. Lab Invest, 2002, 82(6):703-712.
[5] Porokuokka LL, Virtanen HT, Lindén J, et al. Gfra1 underexpression causes Hirschsprung's disease and associated enterocolitis in mice[J]. Cell Mol Gastroenterol Hepatol, 2019, 7(3):655-678.
[6] Chen J, Zhang YC, Huang C, et al. m 6A regulates neurogenesis and neuronal development by modulating histone methyltransferase Ezh2[J]. Genomics Proteomics Bioinformatics, 2019, 17(2):154-168.
[7] Wever I, Wagemans CMRJ, Smidt MP. EZH2 is essential for fate determination in the mammalian isthmic area[J]. Front Mol Neurosci, 2019, 12:76.
[8] Zhou J, Huang S, Wang Z, et al. Targeting EZH2 histone methyltransferase activity alleviates experimental intestinal inflammation[J]. Nat Commun, 2019, 10(1):2427.
[9] Schwarz D, Varum S, Zemke M, et al. Ezh2 is required for neural crest-derived cartilage and bone formation[J]. Development, 2014, 141(4):867-877.
[10] Tien CL, Jones A, Wang H, et al. Snail2/Slug cooperates with Polycomb repressive complex 2(PRC2) to regulate neural crest development[J]. Development, 2015, 142(4):722-731.
[11] Minoux M, Holwerda S, Vitobello A, et al. Gene bivalency at Polycomb domains regulates cranial neural crest positional identity[J]. Science, 2017, 355(6332). pii:eaal2913.
[12] Kim H, Langohr IM, Faisal M, et al. Ablation of Ezh2 in neural crest cells leads to aberrant enteric nervous system development in mice[J]. PLoS One, 2018, 13(8):e0203391.
[13] Mwizerwa O, Das P, Nagy N, et al. Gdnf is mitogenic, neurotrophic, and chemoattractive to enteric neural crest cells in the embryonic colon[J]. Dev Dyn, 2011, 240(6):1402-1411.
[14] Schriemer D, Sribudiani Y, IJpma A, et al. Regulators of gene expression in Enteric Neural Crest Cells are putative Hirschsprung disease genes[J]. Dev Biol, 2016, 416(1):255-265.
[15] Lai FP, Lau ST, Wong JK, et al. Correction of Hirschsprungassociated mutations in human induced pluripotent stem cells via clustered regularly interspaced short palindromic repeats/Cas9, restores neural crest cell function[J]. Gastroenterology, 2017, 153(1):139-153. e8.
[16] Goodman KM, Kjær S, Beuron F, et al. RET recognition of GDNF-GFRα1 ligand by a composite binding site promotes membrane-proximal self-association[J]. Cell Rep, 2014, 8(6):1894-1904.
[17] Meka DP, Müller-Rischart AK, Nidadavolu P, et al. Parkin cooperates with GDNF/RET signaling to prevent dopaminergic neuron degeneration[J]. J Clin Invest, 2015, 125(5):1873-1885.
[18] Wei R, Qiu X, Wang S, et al. NEDL2 is an essential regulator of enteric neural development and GDNF/Ret signaling[J]. Cell Signal, 2015, 27(3):578-586.
[19] 邢晓滨,张卓伯,潘春联,等. GDNF对神经干细胞增殖、分化的影响[J].现代生物医学进展, 2015, 15(33):6583-6586.
[20] Wu W, Lu L, Xu W, et al. Whole exome sequencing identifies a novel pathogenic RET variant in Hirschsprung disease[J]. Front Genet, 2019, 9:752.
[21] Yang D, Yang J, Li S, et al. Effects of RET, NRG1 and NRG3 polymorphisms in a Chinese population with Hirschsprung disease[J]. Sci Rep, 2017, 7:43222.
[22] Kawano T, Hosomichi K, Inoue I, et al. Identification of a novel variant of the RET proto-oncogene in a novel family with Hirschsprung's disease[J]. Pediatr Surg Int, 2017, 33(10):1041-1046.
[23] Borghini S, Bocciardi R, Bonardi G, et al. Hirschsprung associated GDNF mutations do not prevent RET activation[J]. Eur J Hum Genet, 2002, 10(3):183-187.
[24] Angrist M, Jing S, Bolk S, et al. Human GFRA1:cloning, mapping, genomic structure, and evaluation as a candidate gene for Hirschsprung disease susceptibility[J]. Genomics, 1998, 48(3):354-362.
[25] Xu M, Li Y, Du J, et al. PAX3 promotes cell migration and CXCR4 gene expression in neural crest cells[J]. J Mol Neurosci, 2018, 64(1):1-8.
[26] Sun Rhodes LS, Merzdorf CS. The zic1 gene is expressed in chick somites but not in migratory neural crest[J]. Gene Expr Patterns, 2006, 6(5):539-545.
[27] Sham MH, Lui VC, Fu M, et al. SOX10 is abnormally expressed in aganglionic bowel of Hirschsprung's disease infants[J]. Gut, 2001, 49(2):220-226.


PDF(1462 KB)
HTML

Accesses

Citation

Detail

段落导航
相关文章

/