Decreased MiR-200a/141 Suppress Cell Migration and Proliferation by Targeting PTEN in Hirschsprung's Disease

Cellular Physiology and Biochemistry

关键词: HSCR;    3';    -UTR;    Neural crest cell;    Neural development;    Pediatric;   

DOI  :  10.1159/000363021

学科分类：分子生物学，细胞生物学和基因

来源: S Karger AG

【 摘 要 】

Background/Aims Hirschsprung's disease (HSCR) is a genetic disorder of neural crest development. In this study, we investigated whether and how miR-200a and miR-141, belonging to miR-200 family, were involved in the pathogenesis of HSCR. Methods Quantitative real time PCR and Western blot were used to detect the levels of miRNA, mRNAs, and proteins in colon tissues from 88 HSCR patients and 75 controls. The direct regulation of specific mRNA by miRNAs was validated by dual-luciferase reporter assay and RNA interference in cell lines. Transwell assays, CCK8 assay, and flow cytometry were inplemented to measure viability and activities of human 293T and SH-SY5Y cells, respectively. Results Aberrant suppression of miR-200a was observed in colon tissues of HSCR patients. A decreased level of miR-200a and miR-141 correlated with increased levels of PTEN mRNA and protein. The Dual-Luciferase reporter gene assay demonstrated that miR-200a and miR-141 binded directly to 3'UTR of PTEN and resulting in the inhibition of PTEN. The reductions in miR-200a and miR-141 inhibited migration and proliferation of 293T and SH-SY5Y cells through up-regulating the expression of PTEN. Moreover, knocking-down of PTEN rescued the extent of suppressed cell migration and proliferation induced by miR-200a and miR-141. Conclusions The miR-200 family may play a crucial role in the pathogenesis of HSCR by co-regulating PTEN.

【 授权许可】

CC BY-NC-ND