Xiang Luping,Zhai Wei,Xu Shiyu,et al.Mechanism of MEG3 in the pathogenesis of Hirschsprung’s disease through TGF-β mediated regulation of EMT in ENCCs[J].Journal of Clinical Pediatric Surgery,,():852-858.[doi:10.3760/cma.j.cn101785-202510002]
Mechanism of MEG3 in the pathogenesis of Hirschsprung’s disease through TGF-β mediated regulation of EMT in ENCCs
- Keywords:
- Hirschsprung Disease; Long Non-Coding Rna; Epithelial-Mesenchymal Interaction; Enteric Nervous System
- Abstract:
- Objective To investigate the mechanism of MEG3 in regulating the TGF-β/Rac1/JNK pathway to influence the EMT of ENCCs in the pathogenesis of HSCR.Methods Western blot and qRT-PCR were used to detect the expression levels of TGF-β in the stenotic and normal anastomotic segments of the intestine from 40 children with HSCR.Dual-luciferase assays were used to detect the binding and targeted regulatory relationship between MEG3 and TGF-β.ENCCs were extracted from fetal rat intestine,and the expression of EMT markers were detected.Different groups of MEG3 cell models were constructed,and the TGF-β/Rac1/JNK pathway and EMT in each group were detected.CCK-8,cell scratch,and Transwell migration assays were used to observe changes in cell proliferation and migration ability.Results The expression level of TGF-β(0.13±0.01) in the stenotic segments was significantly lower than that in the normal anastomotic segments.Immunofluorescence staining of E-cadherin and N-cadherin in ENCCs were positive.Dual-luciferase reporter assays confirmed that TGF-β is a target gene of MEG3.In MEG3 cell models,compared with the low-expression group and blank control group,the high-expression group showed significantly increased expression of TGF-β(1.31± 0.18),Rac1(1.86±0.24),JNK(1.53±0.03),N-cadherin(0.97±0.10) and Vimentin(1.05±0.12),significantly decreased expression of E-cadherin(0.31±0.23),cell proliferation and migration abilities were both significantly enhanced.The above results were statistically significant( P<0.05).Conclusions High expression of MEG3 can upregulate the TGF-β/Rac1/JNK pathway,promote the EMT process and cell migration and proliferation of ENCCs.Abnormal EMT of ENCCs may be involved in the pathogenesis of HSCR.
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Memo
收稿日期:2025-10-14。
基金项目:遵义市科技计划项目（遵市科合HZ字2025-325号）
通讯作者:刘远梅,Email:yuanmei116@aliyun.com