Comparative transcriptomic analysis of vascular endothelial cells after hypoxia/re-oxygenation induction based on microarray technology

Abstract

Objective: To provide comprehensive data to understand mechanisms of vascular endothelial cell (VEC) response to hypoxia/re-oxygenation. Methods: Human umbilical vein endothelial cells (HUVECs) were employed to construct hypoxia/re-oxygenation-induced VEC transcriptome profiling. Cells incubated under 5% O 2 , 5% CO 2 , and 90% N 2 for 3 h followed by 95% air and 5% CO 2 for 1 h were used in the hypoxia/re-oxygenation group. Those incubated only under 95% air and 5% CO 2 were used in the normoxia control group. Results: By using a well-established microarray chip consisting of 58 339 probes, the study identified 372 differentially expressed genes. While part of the genes are known to be VEC hypoxia/re-oxygenation-related, serving as a good control, a large number of genes related to VEC hypoxia/re-oxygenation were identified for the first time. Through bioinformatic analysis of these genes, we identified that multiple pathways were involved in the reaction. Subsequently, we applied real-time polymerase chain reaction (PCR) and western blot techniques to validate the microarray data. It was found that the expression of apoptosis-related proteins, like pleckstrin homology-like domain family A member 1 (PHLDA1), was also consistently up-regulated in the hypoxia/re-oxygenation group. STRING analysis found that significantly differentially expressed genes SLC38A3, SLC5A5, Lnc-SLC36A4-1, and Lnc-PLEKHJ1-1 may have physical or/and functional protein-protein interactions with PHLDA1. Conclusions: The data from this study have built a foundation to develop many hypotheses to further explore the hypoxia/re-oxygenation mechanisms, an area with great clinical significance for multiple diseases.

Differential angiogenesis of bone and muscle endothelium in aging and inflammatory processes

“…We found that the genes differentially expressed between mEC and bEC were not shared with those differentially expressed between OA grade 2 and grade 3 ECs. Furthermore, when we compared the transcriptomic profile of OA ECs with that of healthy HUVECs 23 , we did not find possible signatures induced by OA in both ECs as compared to a healthy endothelium. Taking these considerations together, we can suppose that the differential expression observed between OA bECs and mECs is mainly attributable to the different origin of endothelial cells, as widely reported in the literature for other healthy tissue types and organisms 2 , 4 .…”