先天性巨结肠中miR-939靶向调控SOX4表达抑制肠神经嵴前体细胞功能表型的实验研究  

Experimental study of miR-939 regulating the phenotypic features of ENCCs by targeting SOX4 in Hirschsprung's disease

作　　者：田东浩 许文耀 余辉[1] 杨薇粒 郑百俊[1] 高亚[1] 潘伟康[1] Tian Donghao;Xu Wenyao;Yu Hui;Yang Weili;Zheng Baijun;Gao Ya;Pan Weikang(Department of Pediatric Surgery,Second affiliated Hospital,Xi'an Jiaotong University,Xi'an 710000,China)

机构地区：[1]西安交通大学第二附属医院小儿外科,陕西省西安市710000

出　　处：《临床小儿外科杂志》2021年第3期222-229,共8页Journal of Clinical Pediatric Surgery

基　　金：国家自然科学基金(编号:NSFC81701501,NSFC82071692,NSFC81770513);陕西省科技新星(编号:2019KJXX-044);陕西省自然科学基金(编号:2020JM-407)。

摘　　要：目的检测MicroRNA-939(miR-939)和性别决定区相关HMG族盒蛋白4(Sry-related HMG box 4,SOX4)在先天性巨结肠(Hirschsprung's disease,HSCR)中的表达水平,评估miR-939在SOX4调控肠神经嵴前体细胞(enteric neural crest cells,ENCCs)增殖、凋亡和迁移过程中的作用,并探讨miR-939/SOX4在HSCR干预治疗方面的潜在价值。方法采用RT-PCR、Western blot检测HSCR和肠套叠患者肠组织中miR-939、SOX4表达水平;从常规孕鼠肠管取材,体外培养ENCCs,通过Nestin、GFAP双免疫荧光染色鉴定。使用miR-939模拟物、SOX4-siRNA干预,并分别通过CCK8法、流式细胞仪和Transwell小室检测ENCCs增殖、凋亡和迁移。结果与肠套叠组患者相比,HSCR患者肠组织中miR-939的表达水平显著提高,而SOX4表达水平显著降低。miR-939模拟物可显著抑制ENCCs中SOX4的表达水平;上调miR-939和下调SOX4均能够显著降低ENCCs增殖和迁移,并促进细胞凋亡。结论HSCR患者肠组织miR-939、SOX4均表达异常,负向调控ENCCs增殖、凋亡和迁移可能参与HSCR发病过程,靶向miR-939/SOX4的干预策略可为后续HSCR干预治疗提供潜在的治疗参考。Objective To detect the expression levels of miR-939 and SOX4 in Hirschsprung's disease(HSCR),evaluate the role of miR-939 in SOX4 regulating enteric neural crest cells(ENCCs)and explore the potential value of miR-939/SOX4 in the intervention of HSCR.Methods Reverse transcription-polymerase chain reaction(RT-PCR)and Western blot were employed for measuring the expression levels of miR-939 and SOX4 in intestinal tissues of HSCR children with intussusception.ENCCs from pregnant Sprague Dawley(SD)rats were routinely cultured and identified by Nestin/GFAP immunofluorescent staining.And miR-939 mimics and SOX4 siRNA were applied to ENCCs and cellular proliferation,apoptosis and migration detected by CCK8,flow cytometry and Transwell chamber assay.Results As compared with intussusception group,the expression level of miR-939 significantly increased in HSCR group while the expression level of SOX4 markedly declined.And miR-939 mimics significantly inhibited the expression of SOX4 in ENCCs.The up-regulation of miR-939 and the down-regulation of SOX4 significantly reduced the proliferation and migration of ENCCs,and promoted apoptosis of ENCCs.Conclusion Conclusion The expression of miR-939/SOX4 is abnormal in HSCR children.A negative regulation of the proliferation,apoptosis and migration of ENCCs may be involved in the pathogenesis of HSCR.Targeted intervention of miR-939/SOX4 signaling pathway provides potential therapeutic targets for subsequent intervention of HSCR.

关 键 词：先天性巨结肠/病因学 肠神经系统 细胞增值 

分 类 号：R574[医药卫生—消化系统] R393[医药卫生—内科学]