RNA-seq-based Technique to Study the Mechanisms Involved in IL-17A Stimulation of Endothelial Cells Leading to Senescence

doi:10.13523/j.cb.2301060

China Biotechnology

2023, Vol. 43

Issue (12): 134-138 DOI: 10.13523/j.cb.2301060

RNA-seq-based Technique to Study the Mechanisms Involved in IL-17A Stimulation of Endothelial Cells Leading to Senescence

CHEN Si-jia,LI Qing(

)

Department of Laboratory Medicine, The First Affiliated Hospital of the University of Science and Technology of China, Hefei 230001, China

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Abstract

Objective: To study the differential gene expression in senescent endothelial cells after IL-17 induction and to explore possible mechanisms of action of IL-17-induced endothelial cell senescence. Methods: Unstimulated human umbilical vein epithelial cells (HUVEC) and IL-17-stimulated HUVEC were subjected to senescence index assay and RNA-seq sequencing, and the differential genes were analyzed by KEGG enrichment, and the differential genes were screened for qRT-PCR validation. Results: Cellular senescence occurred in IL-17-stimulated HUVEC compared to unstimulated HUVEC. After cluster analysis, SMAD6 was selected for qRT-PCR validation. The results of the analysis were consistent with the trend of the sequencing results. Conclusions: IL-17 can activate or inhibit a number of senescence-related genes and pathways that lead to endothelial cell senescence.

Key words： Vascular endothelial cells Cellular senescence RNA-seq IL-17

Received: 10 October 2023 Published: 16 January 2024

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Si-jia CHEN, Qing LI. RNA-seq-based Technique to Study the Mechanisms Involved in IL-17A Stimulation of Endothelial Cells Leading to Senescence. China Biotechnology, 2023, 43(12): 134-138.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2301060 OR https://manu60.magtech.com.cn/biotech/Y2023/V43/I12/134

Fig.1 Effect of IL-17 stimulation on HUVEC senescence indicators qRT-PCR was used to determine p53, p21, and p16 in two groups of HUVEC (n = 3);Values expressed as mean ± SD. ^** P<0.01, ^*** P<0.001

Fig.2 Distribution map of DEG A. Number of DEGs with increased and decreased expression in IL-17-stimulated endothelial cells compared to unstimulated endothelial cells B. Hierarchical cluster analysis of differentially expressed genes

Fig.3 KEGG significance bubble plot Significantly enriched for KEGG pathways. x-axis shows concentration scores, y-axis shows enrichment for KEGG pathways. The size of the circle indicates the number of genes and the color indicates the P-value

Fig.4 Validation of aging-related differential genes A. Volcano plot showing differential genes. y-axis shows values of - log₁₀ (P) and x-axis shows values of log₂-fold change. Genes with significantly increased and decreased expression are indicated by red and green dots, respectively, and blue dots indicate genes with no significant change B. RNA expression of SMAD6 by qRT-PCR. Values expressed as mean ± SD. ^* P< 0.05


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