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Coronin 1C negatively regulates cell-matrix adhesion and motility of intestinal epithelial cells.

ABSTRACT: Coronins, WD-repeat actin-binding proteins, are known to regulate cell motility by coordinating actin filament turnover in lamellipodia of migrating cell. Here we report a novel mechanism of Coronin 1C-mediated cell motility that involves regulation of cell-matrix adhesion. RNAi silencing of Coronin 1C in intestinal epithelial cells enhanced cell migration and modulated lamellipodia dynamics by increasing the persistence of lamellipodial protrusion. Coronin 1C-depleted cells showed increased cell-matrix adhesions and enhanced cell spreading compared to control cells, while over-expression of Coronin 1C antagonized cell adhesion and spreading. Enhanced cell-matrix adhesion of coronin-deficient cells correlated with hyperphosphorylation of focal adhesion kinase (FAK) and paxillin, and an increase in number of focal adhesions and their redistribution at the cell periphery. siRNA depletion of FAK in coronin-deficient cells rescued the effects of Coronin 1C depletion on motility, cell-matrix adhesion, and spreading. Thus, our findings provide the first evidence that Coronin 1C negatively regulates epithelial cell migration via FAK-mediated inhibition of cell-matrix adhesion.

SUBMITTER: Samarin SN

PROVIDER: S-EPMC2812594 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Coronin 1C negatively regulates cell-matrix adhesion and motility of intestinal epithelial cells.

Samarin Stanislav N SN Koch Stefan S Ivanov Andrei I AI Parkos Charles A CA Nusrat Asma A

Biochemical and biophysical research communications 20091112 1

Coronins, WD-repeat actin-binding proteins, are known to regulate cell motility by coordinating actin filament turnover in lamellipodia of migrating cell. Here we report a novel mechanism of Coronin 1C-mediated cell motility that involves regulation of cell-matrix adhesion. RNAi silencing of Coronin 1C in intestinal epithelial cells enhanced cell migration and modulated lamellipodia dynamics by increasing the persistence of lamellipodial protrusion. Coronin 1C-depleted cells showed increased cel ...[more]

PMID: 19913511

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Project description:Successful embryo implantation requires a competent embryo, a receptive endometrium and synchronized communication between them. The selection of embryos with the highest implantation potential remains a challenge in the field of assisted reproductive technology. Moreover, little is known about the precise molecular mechanisms underlying embryo‑endometrium crosstalk. MicroRNAs (miRNAs/miRs) have been detected in the spent embryo culture medium (SCM); however, their functions at the preimplantation stage remain unclear. In the present study, human SCM samples were collected during in vitro fertilization/intracytoplasmic sperm injection‑embryo transfer and divided into implanted and not‑implanted groups according to the clinical pregnancy outcomes. Total RNA was extracted and six miRNAs (miR‑372‑3p, miR‑373‑3p, miR‑516b‑5p, miR‑517a‑3p, miR‑519d‑3p and miR‑520a‑3p) were selected for reverse transcription‑quantitative PCR (RT‑qPCR) analysis. The results revealed that miR‑372‑3p and miR‑519d‑3p were markedly increased in SCM from blastocysts that failed to implant compared with in blastocysts that implanted. The receiver operating characteristic curve analysis revealed that miR‑519d‑3p was superior to miR‑372‑3p in predicting pregnancy outcomes. In vitro miRNA uptake and cell adhesion assays were performed to determine whether miR‑519d‑3p could be taken up by endometrial epithelial cells and to examine the biological roles of miR‑519d‑3p after internalization. Potential targets of miR‑519d‑3p were verified using a dual‑luciferase reporter system. The results demonstrated that miR‑519d‑3p was taken up by human endometrial epithelial cells and that it may inhibit embryo adhesion by targeting HIF1α. Using RT‑qPCR, western blot analysis and flow cytometry assay, HIF1α was shown to inhibit the biosynthesis of fucosyltransferase 7 and sialyl‑Lewis X (sLex), a cell‑surface oligosaccharide that serves an important role in embryonic apposition and adhesion. In addition, a mouse model was established and the results suggested that miR‑519d‑3p overexpression hampered embryo implantation in vivo. Taken together, miRNAs in SCM may serve as novel biomarkers for embryo quality. Furthermore, miR‑519d‑3p was shown to mediate embryo‑endometrium crosstalk and to negatively regulate embryo implantation by targeting HIF1α/FUT7/sLex pathway.

Dataset Information

Coronin 1C negatively regulates cell-matrix adhesion and motility of intestinal epithelial cells.

Publications

Coronin 1C negatively regulates cell-matrix adhesion and motility of intestinal epithelial cells.

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