E-MTAB-1173 - Transcription profiling by array of human HUVEC, HCAEC and MM6 cell lines for mechanistic investigation of the impact of cigarette smoke on adhesion between monocytes and endothelial cells in vitro
Submitted on 14 June 2012, released on 24 April 2014, last updated on 12 September 2014
The adhesion of monocytic cells to the “dysfunctional” endothelium constitutes a critical step in the initiation of atherosclerotic plaque formation. Cigarette smoke (CS) has been shown to contribute to the monocyte-endothelial adhesion process. Yet, the complex mechanisms underlying this event remain to be discovered. In the systemic compartment, monocytes and endothelial cells are exposed primarily to soluble constituents originating from the inhaled CS and absorbed through the lung alveolar epithelium and the adjacent microvascular endothelium. Considering this rationale, we developed an in vitro adhesion assay, intending to mimic the in vivo situation, to deeply investigate the impact of CS on the adhesion of monocytic cell to the endothelium. Using a transcriptomics approach followed by confirmation experiments, we were able to identify a key mechanism by which aqueous extract of CS (smoke-bubbled Phosphate Buffered Saline, sbPBS) promotes the adhesion of monocytic Mono Mac 6 (MM6) cells to the human umbilical vein endothelial cells (HUVECs). While CS-derived constituents directly provoke a strong oxidative stress response in both cell types, the induced expression of E-selectin, VCAM-1 and ICAM-1 adhesion molecules, responsible for the binding of MM6 cells to HUVECs, occurs through a proinflammatory paracrine effect. We demonstrate that this effect is mainly driven by TNF? produced by MM6 cells exposed to sbPBS. In conclusion, the development of our in vitro adhesion assay, intending to mimic the in vivo conditions, enabled to show that the adhesion of monocytic cells to endothelial cells is promoted through an indirect effect of the sbPBS.
transcription profiling by array, cell type comparison, compound treatment, dose response, in vitro, time series
Mechanism of an indirect effect of aqueous cigarette smoke extract on the adhesion of monocytic cells to endothelial cells in an in vitro assay revealed by transcriptomics analysis. Poussin C, Gallitz I, Schlage WK, Steffen Y, Stolle K, Lebrun S, Hoeng J, Peitsch MC, Lietz M. Toxicol In Vitro (2014), Europe PMC 24747719