Differential Expression And Preliminary Analysis Of Hepatocellular Carcinoma Gene Based On High-throughput Sequencing

Objective: High-throughput sequencing technology was used to sequence and analyze the differentially expressed genes in HBV-related hepatocellular carcinoma tissues and paired adjacent tissues,and bioinformatics was used to analyze differentially expressed genes,revealing the possible key regulatory pathways and differentially expressed genes in the development of hepatocellular carcinoma,laying a foundation for further research.Method:1.Analysis of differentially expressed RNA in hepatocellular carcinoma.HBV-related hepatocellular carcinoma(HCC)and its adjacent tissues were sequenced by high-throughput sequencing(3 cases each).The expression of differentially expressed genes was calculated,and the differentially expressed genes were screened to construct the differentially expressed genes of HCC.2.Bioinformatics was used to analyze differentially expressed RNA.GO function enrichment analysis,KEGG signal pathway enrichment analysis and KO analysis of differentially expressed genes were carried out to reveal thepossible differentially expressed functions of differentially expressed genes and the metabolic pathways and signal pathways in which differentially expressed genes are mainly involved.Combined with the results of enrichment analysis,key regulatory pathways and differentially expressed genes were screened.3.Expanding sample size verification.HBV-related hepatocellular carcinoma tissues and their paired paracancerous tissues(8 cases each)were examined by real-time fluorescent quantitative PCR and immunohistochemical techniques at gene and protein levels to determine whether their expression was consistent with high-throughput sequencing results,which laid a foundation for further research.Result:1.A total of 2386 differentially expressed genes were screened by high-throughput sequencing analysis.Among them,119 were up-regulated and1267 were down-regulated in paracancerous group compared with hepatocellular carcinoma group.2.Functional enrichment analysis of differentially expressed genes GO showed that the biological processes GO Term involved in differentially expressed genes were: GO:0046395 carboxylic acid catabolic process,GO:0032787monocarboxylic acid metabolic process and GO:1901605 alpha-amino acid metabolic process.As a cell component,differentially expressed genes are mainly concentrated in GO Term,such as GO:0070062 extracellular exosome,GO:0044444 cytoplasmic part and GO:0044459 plasma membrane part;differentially expressed genes are mainly concentrated in GO:0016614oxidoreductase activity,acting on CH-OH group of donor.GO Te,such as s,GO: 0042802 identical protein binding,GO: 0043168 anion binding,GO:0050662 coenzyme binding and GO: 0008028 monocarboxylic acidtransmembrane Porter activity RM.Enrichment analysis of KEGG signaling pathway revealed that differentially expressed genes mainly concentrated in Valine,leucine and isoleucine degradation,Tyrosine metabolism(tyrosine metabolism),Tryptophan metabolism,Retinol metabolism,Propanoate metabolism,PPAR signaling P.Athway,Peroxisome,Metabolism of xenobiotics by cytochrome P450,Glycine,serine and threonine metabolism,Fatty acid degradation,Drug metabolism-other enzymes.Other enzymes,Drug metabolism-cytochrome P450,Complement and coagulation cascades,Cell cycle,Carbon metabolism,Caffeine metabolism,Butanoate metabolism,Bile secretion,beta-Alanine M Etabolism,Arginine and proline metabolism and other 20 signaling pathways.KO analysis revealed that SFN,CCNB1 and CDK1 were up-and-down in the cell cycle pathway and significantly up-regulated(HCC tissues relative to paired adjacent tissues).3.The expression levels of SFN,CCNB1 and CDK1 genes and proteins were verified by real-time fluorescence quantitative PCR and immunohistochemistry.Compared with the adjacent tissues,the expression levels of SFN,CCNB1 and CDK1 genes and proteins in hepatocellular carcinoma tissues were up-regulated,with significant difference(P < 0.05).Conclusion:1.A total of 2386 differentially expressed genes were screened by high-throughput sequencing,of which 1119 were up-regulated and 1267 were down-regulated in paracancerous tissues compared with hepatocellular carcinoma tissues.2.SFN,CCNB1 and CDK1 are highly expressed in hepatocellular carcinoma tissues and are enriched in cell cycle pathways.