| 摘要: |
| 目的:探究miR-429 调控NRG1 对先天性巨结肠( HSCR) 神经嵴细胞增殖和迁移的影响。方法:qRT-PCR 检测35 例
HSCR 患儿和35 例非HSCR 肠套叠或腹股沟疝患儿结肠组织中miR-429 和NRG1 mRNA 表达水平。培养293T 细胞和SH-SY5Y
细胞,依次分为control 组、miR-429 NC 组、miR-429 组、miR-429+OE 组和miR-429+OE-NRG1 组。通过qRT-PCR 和Western blot
检测miR-429 和NRG1 表达鉴定转染效率;CCK-8、EdU、Transwell 和Western blot 检测细胞增殖、迁移及相关蛋白表达;双荧光素
酶实验证实miR-429 和NRG1 之间存在相互作用。结果:HSCR 患儿结肠组织中miR-429 表达下调,NRG1 表达上调。与
miR-429 NC 组比较,miR-429 组293T、SH-SY5Y 细胞中miR-429 表达、OD 值、EdU 阳性细胞率、CDK6 蛋白表达、c-Myc 蛋白表
达、迁移细胞数量、MMP-9 蛋白表达、Slug 蛋白表达、Snail 蛋白表达均升高,NRG1 表达降低(P<0. 05);与miR-429+OE 组比较,
miR-429+OE-NRG1 组293T、SH-SY5Y 细胞中NRG1 表达升高,OD 值、EdU 阳性细胞率、CDK6 蛋白表达、c-Myc 蛋白表达、迁移
细胞数量、MMP-9 蛋白表达、Slug 蛋白表达、Snail 蛋白表达均降低( P<0. 05)。结论:miR-429 能靶向调控NRG1 表达。上调
miR-429 通过抑制NRG1 表达,促进HSCR 肠神经嵴细胞增殖和迁移。 |
| 关键词: miR-429 NRG1 先天性巨结肠 神经嵴细胞 |
| DOI：10.13407/j.cnki.jpp.1672-108X.2025.12.005 |
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| 基金项目:河北省医学科学研究重点课题计划项目,编号20240471。 |
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| Clinical Research of miR-429 in Promoting Proliferation and Migration of Enteric Neural Crest Cells inHirschsprung’s Disease by Targeting NRG1 |
| Li Xuequn, Zhang Haisheng |
| (Tangshan Maternal and Child Health Hospital, Hebei Tangshan 063000, China) |
| Abstract: |
| Objective: To investigate the effects of miR-429 in promoting proliferation and migration of enteric neural crest cells in
Hirschsprung’s disease (HSCR) by targeting NRG1. Methods: The expression levels of miR-429 and NRG1 mRNA were detected by
qRT-PCR in colonic tissues from 35 cases of children with HSCR and 35 cases of intussusception or inguinal hernia in children without
HSCR. 293T cells and SH-SY5Y cells were cultured and divided into the control group, miR-429 NC group, miR-429 group, miR-429
+OE group, and miR-429+OE-NRG1 group. qRT-PCR and Western blot were used to determine the transfection efficiency by detecting
the expression of miR-429 and NRG1. Cell proliferation, migration capacity, and protein expression were assessed by CCK-8, EdU,
Transwell, and Western blot. The interaction between miR-429 and NRG1 was confirmed by dual-luciferase reporter assay. Results:
Downregulated miR-429 and upregulated NRG1 were observed in HSCR tissues. Compared with the miR-429 NC group, the miR-429
group showed a significant increase in miR-429 expression, OD value, EdU positive cell rate, CDK6 protein, c-Myc protein, migration
cell number, MMP-9 protein, Slug protein, and Snail protein in 293T and SH-SY5Y cells, while a decrease in NRG1 (P <0. 05).
Compared with the miR-429+OE group, the miR-429+OE-NRG1 group exhibited a significant increase in NRG1 expression, while a
decrease in OD value, EdU positive cell rate, CDK6 protein, c-Myc protein, migration cell number, MMP-9 protein, Slug protein, and
Snail protein expression in 293T and SH-SY5Y cells (P<0. 05). The expression of NRG1 was regulated by miR-429 through targeted
regulation (P<0. 05). Conclusion: miR-429 can target and regulate the expression of NRG1. Upregulation of miR-429 promotes the
proliferation and migration of enteric neural crest cells in HSCR by inhibiting the expression of NRG1. |
| Key words: miR-429 NRG1 Hirschsprung’s disease neural crest cells |
