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Published on: April 26, 2019
Aberrant reduction of MiR-141 increased CD47/CUL3 in Hirschsprung's disease
Weibing Tang1, Jingjing Qin, Junwei Tang
1State Key Laboratory of Reproductive Medicine, Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, China.
Background:
MiR-141 has been confirmed to be associated with various human diseases. However, whether miR-141 is involved in the pathogenesis of Hirschsprung's disease (HSCR) remains unknown. Here, we design the experiment to reveal the relationship between miR-141 and HSCR.
Methods:
Quantitative real-time PCR and Western blot were used to detect the expression levels of miR-141 and its potential genes in 70 tissues of HSCR compared with 60 controls. Bisulfite sequencing PCR (BSP) assay was applied to explain the possible mechanism of the aberrant expression level of miR-141. We employed a dual-luciferase reporter assay to validate the regulation relation between miR-141 and CD47/CUL3. Cell migration, proliferation, apoptosis, and cell cycle progression were examined by transwell assay, MTT assay, and flow cytometry, respectively.
Results:
MiR-141 was down-regulated whereas CD47 and CUL3 expression was increased in colon tissues from patients with HSCR compared with control group, The increased level of CD47 and CUL3 induced by miR-141 reduced proliferation and migration of 293T and SH-SY5Y cells. Furthermore, this suppression was reversed by reducing of CD47 and CUL3. Hypermethylation of a CpG Island in the promoter region of miR-141 gene was confirmed in HSCR tissues.
Conclusion:
Aberrant reduction of miR-141 may play an important role in the pathogenesis of HSCR with the inhibiting affection on cell migration and proliferation abilities. The present study demonstrates for the first time the role of miR-141 and its target genes in the occurrence of HSCR, and provides us a new direction for the study of the pathogenesis of Hirschsprung's disease.
Insights
MicroRNA-141 (miR-141) reduction is linked to Hirschsprung's disease (HSCR) pathogenesis. Its dysregulation affects cell migration and proliferation, offering new insights into HSCR development.
Area of Science:
- Genetics
- Developmental Biology
- Molecular Biology
Background:
- MicroRNA-141 (miR-141) is implicated in various human diseases.
- The role of miR-141 in the pathogenesis of Hirschsprung's disease (HSCR) was previously unknown.
Purpose of the Study:
- To investigate the relationship between miR-141 and Hirschsprung's disease (HSCR).
- To elucidate the molecular mechanisms underlying miR-141 dysregulation in HSCR.
Main Methods:
- Quantitative real-time PCR and Western blot to assess miR-141, CD47, and CUL3 expression.
- Bisulfite sequencing PCR (BSP) to analyze promoter methylation.
- Dual-luciferase reporter assay to confirm miR-141 targeting of CD47/CUL3.
- Cellular assays (Transwell, MTT, flow cytometry) to evaluate cell behavior.
Main Results:
- MiR-141 was significantly down-regulated in HSCR tissues, while CD47 and CUL3 were up-regulated.
- Increased CD47 and CUL3, induced by miR-141 reduction, inhibited cell migration and proliferation.
- Hypermethylation of the miR-141 promoter region was observed in HSCR tissues.
- Reversal of suppression was achieved by reducing CD47 and CUL3 levels.
Conclusions:
- Down-regulation of miR-141 plays a crucial role in Hirschsprung's disease (HSCR) pathogenesis.
- MiR-141 affects cell migration and proliferation, potentially through its target genes CD47 and CUL3.
- This study provides novel insights into the molecular mechanisms of HSCR and identifies miR-141 as a potential therapeutic target.
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