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Simultaneous Study of the Recruitment of Monocyte Subpopulations Under Flow In Vitro
Published on: November 26, 2018
miR-27a inhibits molecular adhesion between monocytes and human umbilical vein endothelial cells; systemic approach
Farhad Shaikhnia1, Ghasem Ghasempour1, Asghar Mohammadi2
1Clinical Biochemistry Department, Faculty of Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Insights
MicroRNAs miR-27a and miR-194 reduce key adhesion molecules, significantly decreasing monocyte adhesion to endothelial cells and impacting leukocyte diapedesis.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Endothelial cells overexpress adhesion molecules, promoting leukocyte diapedesis and subendothelial molecular events.
- Leukocyte tethering involves specific adhesion molecules like SELE, SELP, and JAM-B.
- MicroRNAs (miRNAs) play crucial roles in regulating gene expression, including adhesion molecule pathways.
Purpose of the Study:
- To investigate the role of miR-194 and miR-27a in regulating adhesion molecule expression in human umbilical vein endothelial cells (HUVECs).
- To determine the effect of these miRNAs on monocyte-endothelial cell adhesion.
- To explore the potential involvement of miR-194 and miR-27a in the leukocyte diapedesis pathway.
Main Methods:
- Prediction of adhesion molecules (SELE, SELP, JAM-B) involved in leukocyte tethering using a gene network analysis.
- Transfection of HUVECs with PEI-miRNA particles carrying miR-194 and miR-27a.
- Quantification of SELE, SELP, and JAM-B gene expression using real-time quantitative polymerase chain reaction (qPCR).
- Assessment of monocyte-endothelial cell adhesion using an adhesion assay kit.
Main Results:
- miR-194 and miR-27a significantly decreased the expression of SELP and JAM-B in HUVECs (P < 0.05).
- Both miRNAs suppressed monocyte adhesion to endothelial cells.
- miR-27a demonstrated a significant inhibition of monocyte-endothelial adhesion (P = 0.0001) by suppressing SELP and JAM-B.
Conclusions:
- miR-194 and miR-27a effectively reduce the expression of SELP and JAM-B in endothelial cells.
- These miRNAs inhibit monocyte adhesion to endothelial cells, suggesting a role in regulating leukocyte diapedesis.
- miR-27a, in particular, shows significant potential in modulating leukocyte diapedesis through SELP and JAM-B suppression.
Objective:
The endothelial cells overexpress the adhesion molecules in the leukocyte diapedesis pathway, developing vessel subendothelial molecular events. In this study, miR-194 and miR-27a were predicted and investigated on the expression of adhesion molecules in HUVEC cells. The SELE, SELP, and JAM-B adhesion molecules involved in the leukocyte tethering were predicted on the GO-enriched gene network. Following transfection of PEI-miRNA particles into HUVEC cells, the SELE, SELP, and JAM-B gene expression levels were evaluated by real-time qPCR. Furthermore, the monocyte-endothelial adhesion was performed using adhesion assay kit.
Results:
In agreement with the prediction results, the cellular data showed that miR-27a and miR-194 decrease significantly the SELP and JAM-B expression levels in HUVECs (P < 0.05). Moreover, both the miRNAs suppressed the monocyte adhesion to endothelial cells. Since the miR-27a inhibited significantly the monocyte-endothelial adhesion (P = 0.0001) through the suppression of SELP and JAM-B thus it might relate to the leukocyte diapedesis pathway.
