Study On Screening Key Genes In Aldosterone-producing Adenoma Based On Transcriptome Sequencing

Objective:Aldosterone-producing adenoma(APA)is a complex disease caused by a variety of factors.Previous research results cannot fully explain the occurrence and development of APA.This research aims to screen out the key genes of APA and analyze the single nucleotide polymorphism(SNP)mutation sites of the key genes,through transcriptome sequencing of clinical tissue samples,and further bioinformatics analyzing combined with public databases,and then verifying the expression level of differentially expressed genes(DEGs).Methods:(1)Transcriptome sequencing was performed on 12 adrenal gland tissue samples from clinically confirmed APA patients.After obtaining DEGs,perform GO and KEGG analysis,and construct a protein-protein interaction(PPI)network to screen the genes of key modules.(2)Get the GSE156931,GSE60042 and GSE8514 data series with keywords in the GEO public database,and then use GEO2 R to analyze and take the intersection to get common DEGs.Perform functional enrichment analysis on common DEGs,and integrate transcriptome sequencing analysis for further screening(3)Tissue samples from APA patients were collected and divided into two groups: adrenal adenoma tissue and normal adrenal tissue adjacent to the tumor.Real-time quantitative PCR(q PCR)experiments were performed to verify the expression of the above-mentioned key genes at the m RNA level.(4)The SNP mutation site of the key gene CYP11B2 was found,annotated based on the results of transcriptome sequencing,and searched through databases and literature.Results:(1)704 DEGs were obtained based on transcriptome sequencing,and then construct a PPI network of key gene modules,and GABBR2,NMU,HCAR2,SSTR1,PTGER3,ADCY8,ADCY2,FPR2,CXCL9,CXCL6,PTGDR2,GHRL,SNAP25 were screened as possible key genes of APA.(2)The 311 DEGs were mined by bioinformatics analysis.Combined with the DEGs obtained by transcriptome sequencing,it was found that CYP11B2 and PNLIP were significantly different,and CYP11B2 was closely related to aldosterone synthesis,which may be the key gene of APA.(3)The 14 key genes previously screened were verified by q PCR,and it was found that the m RNA expression level of CYP11B2 in APA adenoma tissue was significantly higher than that in normal adrenal tissue.(4)In this study,a total of 65 SNP mutation sites in the CYP11B2 gene were detected,of which 6 SNP mutation sites affect the amino acid sequence,and 3 SNP mutation sites c.*5738A>G,c.*5758A>G,c.396-561C>A have not been reported.Conclusion:(1)CYP11B2 was screened out as the key gene of APA,based on transcriptome sequencing and bioinformatics analysis,and was verified by q PCR experiments.(2)A total of 65 SNP mutation sites in the CYP11B2 gene were detected,of which c.*5738A>G,c.*5758A>G,and c.396-561C>A are newly discovered mutation sites,which are worthy of further study.