MicroRNA-15a - cell division cycle 42 signaling pathway in pathogenesis of pediatric inflammatory bowel disease

Abstract

AIMTo determine whether cell division cycle (Cdc)42 is regulated by microRNA (miR)-15a in the development of pediatric inflammatory bowel disease (IBD).METHODSWe cultured 293T cells, used plasmids and performed dual-luciferase assay to determine whether Cdc42 is a miR-15a target gene. We cultured Caco-2 cells, and stimulated them with tumor necrosis factor (TNF)-α. We then employed lentiviruses to alter the expression of miR-15a and Cdc42. We performed quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunofluorescence to determine whether Cdc42 is regulated by miR-15a in Caco-2 cells. Finally, we collected ileocecal tissue by endoscopy from patients and performed qRT-PCR to examine the expression of miR-15a and Cdc42 in pediatric IBD patients.RESULTSTarget Scan and dual-luciferase assay revealed that Cdc42 was a miR-15a target gene. MiR-15a expression increased (P = 0.0038) and Cdc42 expression decreased (P = 0.0013) in cells stimulated with TNF-α, and the expression of the epithelial junction proteins zona occludens (ZO)-1 (P < 0.05) and E-cadherin (P < 0.001) decreased. Cdc42 levels decreased in miR-15a-mimic cells (P < 0.001) and increased in miR-15a inhibitor cells (P < 0.05). ZO-1 and E-cadherin decreased in miR-15a-mimic cells (P < 0.001) but not in the miR-15a inhibitor + TNF-α cells. In Lv-Cdc42 + TNF-α cells, ZO-1 and E-cadherin expression increased compared to the Lv-Cdc42-NC + TNF-α (P < 0.05) or miR-15a-mimic cells (P < 0.05). Fifty-four pediatric IBD patients were included in this study, 21 in the control group, 19 in the Crohn’s disease (CD) active (AC) group, seven in the CD remission (RE) group, and seven in the ulcerative colitis (UC) group. MiR-15a increased and Cdc42 decreased in the CD AC group compared to the control group (P < 0.05). miR-15a decreased and Cdc42 increased in the CD RE group compared to the CD AC group (P < 0.05). miR-15a was positively correlated with the Pediatric Crohn’s disease Activity Index (PCDAI) (P = 0.006), while Cdc42 was negatively correlated with PCDAI (P = 0.0008). Finally, miR-15a expression negatively correlated with Cdc42 in pediatric IBD patients (P = 0.0045).CONCLUSIONMiR-15a negatively regulates epithelial junctions through Cdc42 in Caco-2 cells and pediatric IBD patients.

<p>miR-26b-5p Inhibits the Proliferation, Migration and Invasion of Human Papillary Thyroid Cancer in a β-Catenin-Dependent Manner</p>

“…MicroRNAs are long noncoding RNAs that are involved in vital cellular processes, such as cell division and differentiation. 2,3 MicroRNAs have also been reported to participate in the tumourigenesis and progression of multiple cancers. [4][5][6] However, the function of the same microRNA varies in different kinds of cancers.…”

Cell Division Cycle 2 Protects Neonatal Rats Against Hyperoxia-Induced Bronchopulmonary Dysplasia

“…26 CDC42 was down-regulated in the pediatric inflammatory bowel disease. 27 In this study, the protein and mRNA expressions of CDC2 in lung tissues were markedly decreased in the Model group on days 7 and 14 and Hyperoxia+pcDNA-CDC2 † † ‡ ‡ † ‡ ‡ † † † † † † † † † † † † † † † † † † Fig. 4.…”

“… 26 CDC42 was down-regulated in the pediatric inflammatory bowel disease. 27 In this study, the protein and mRNA expressions of CDC2 in lung tissues were markedly decreased in the Model group on days 7 and 14 and in hyperoxia-induced A549 cells. The results suggest that CDC2 may play important roles in regulating hyperoxia-induced BPD.…”

Rho GTPases as Key Molecular Players within Intestinal Mucosa and GI Diseases

“…In the context of human intestinal inflammation, the expression of CDC42 is decreased in active IBD, and CDC42 has been suggested as a target of MiR-15a for the regulation of epithelial junctions (ZO1 and E-cadherin) in pediatric patients [ 156 ]. In contrast, another study describe the upregulation of ACK1 (Activated CDC42 kinase1) in colitis as well as colorectal dysplasia [ 157 ].…”

“…A recent publication describes the mechanism by which HuR (RNA-binding protein HuR) promotes protein expression of CDC42 within IECs, contributing to epithelial restitution (healing) in ischemia as well as colitis models (DSS), in a mechanism that is dependent on the actin cytoskeleton [ 173 ]. As mentioned above, MiR-15a regulates epithelial junctions via CDC42-dependent mechanisms in pediatric IBD patients [ 156 ].…”