Screening and functional validation of core pathogenic genes in aldosterone-producing adenoma based on integrated transcriptomic and proteomic analysis

doi:10.11958/20260728

Abstract

Abstract:

Objective To identify key pathogenic genes in aldosterone-producing adenoma (APA) and validate their biological functions through integrated transcriptomic and proteomic analysis. Methods Three samples each of APA tissue and control tissue from non-functioning adrenal adenomas were collected, and whole-transcriptome sequencing (RNA-seq) and quantitative proteomics were performed simultaneously. Based on integrated transcriptomic and proteomic analysis, differentially expressed genes and proteins were identified, and core pathways and key molecules were identified through functional enrichment analysis. Furthermore, in vitro cellular experiments were conducted using overexpression and silencing techniques to regulate fatty acid synthase (FASN) expression, and cell proliferation rates, adenosine triphosphate (ATP) levels, and the expression of aldosterone synthase (CYP11B2) protein were assessed. Results A total of 10 705 quantifiable transcripts and 3 288 quantifiable proteins were identified, of which 907 molecules were common to both omics datasets. Differential analysis revealed that 167 molecules were upregulated at both transcriptional and protein levels in APA, whilst 213 molecules were downregulated in a coordinated manner. Functional enrichment analysis indicated that the synergistically up-regulated molecules were significantly enriched in lipid metabolism-related pathways, such as fatty acid biosynthesis, the peroxisome proliferator-activated receptor (PPAR) signalling pathway and cholesterol metabolism, with the fatty acid biosynthesis pathway showing the highest level of enrichment. Further screening identified 10 key molecules commonly enriched across these core pathways; among these, FASN—the rate-limiting enzyme in fatty acid synthesis—was significantly upregulated in APA at both the messenger RNA (mRNA) and protein levels. Overexpression of FASN significantly promoted cell proliferation (P<0.01), reduced ATP production (P<0.01), and upregulated CYP11B2 protein expression (P<0.01). In contrast, silencing of FASN produced the opposite results (P<0.01). Conclusion As a key rate-limiting enzyme in the fatty acid synthesis pathway, FASN promotes tumour cell proliferation and the expression of a key enzyme involve in aldosterone synthesis, and may be involved in the pathogenesis of APA.

Key words: hyperaldosteronism, transcriptome, proteome, fatty acid synthase, cell proliferation, fatty acid metabolism

CLC Number:

R318.15，R736

Figures/Tables 5

Fig.1 Quality test for integrated transcriptomic and proteomic analysis of APA and NFALs

Fig.2 KEGG pathway and GO function enrichment analysis of upregulated genes in transcriptomic and proteomic analysis

Fig.3 Proliferation curves of transfected cells of each group

Fig.4 ATP content of transfected cells of each group

Fig.5 Expression levels of FASN, CYP11B2 in transfected cells of each group

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