Comparative transcriptomic analysis of vascular endothelial cells after hypoxia/re-oxygenation induction based on microarray technology.
- Author:
Jia XU
1
;
Jiu-Kun JIANG
1
;
Xiao-Lin LI
1
;
Xiao-Peng YU
2
;
Ying-Ge XU
1
;
Yuan-Qiang LU
1
Author Information
- Publication Type:Journal Article
- Keywords: Human umbilical vein endothelial cells (HUVECs); Hypoxia; Re-oxygenation; Microarray; Pleckstrin homology-like domain family A member 1 (PHLDA1); Long non-coding RNA (lncRNA)
- MeSH: Cell Hypoxia; Cells, Cultured; Computational Biology; Human Umbilical Vein Endothelial Cells/metabolism*; Humans; Microarray Analysis/methods*; Transcription Factors/genetics*; Transcriptome
- From: Journal of Zhejiang University. Science. B 2020;21(4):291-304
- CountryChina
- Language:English
-
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% O2, 5% CO2, and 90% N2 for 3 h followed by 95% air and 5% CO2 for 1 h were used in the hypoxia/re-oxygenation group. Those incubated only under 95% air and 5% CO2 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.
