Screening Of Differential Expression Profiles Of CircRNAs In Cholangiocarcinoma And Preliminary Study On The Role Of Hsa_circRNA₀00585 In The Pathogenesis Of Cholangiocarcinoma

Part ?Screening and bioinformatics analysis of differential expression profiles of circular RNA in tissues of cholangiocarcinomaand adjacent tissuesOBJECTIVE: To detect the expression profiles of circulatory RNA?circRNAs?in cholangiocarcinomas?CCA?and paracancerous tissues,and to screen differentially expressed circRNAs in cancer tissues and adjacent tissues.Bioinformatics analysis was used to predict differentially expressed circRNAs target genes.Biological functions and related pathways to further explore the molecular mechanisms of cholangiocarcinoma.METHODS: The expression profiles of circRNAs in three groups of matched cholangiocarcinoma tissues and adjacent normal tissues were sequenced by gene chip.The differential expression of circRNAs was screened.The differentially expressed circRNAs were analyzed by bioinformatics to predict the related functions and signaling pathways of downstream target genes.The Arraystar software predicts the downstream target miRNA bound by the selected hsa_circRNA₀0058515 and details the hsa_circRNA₀0058515/miRNA interaction.RESULTS: A total of 13253 circRNAs were detected by gene chip.697 differentially expressed circRNAs were screened by P<0.05.Among them,387 circRNAs were up-regulated and 310 circRNAs were down-regulated.According to the fold change?FC?>1.5 and P <0.05 as screening criteria,221 differentially expressed circRNAs were screened out,of which 117 circRNAs were up-regulated and 104 circRNAs were down-regulated.GO enrichment analysis found that cholangiocarcinoma tissue up-regulated expression of circRNAs host genes may be involved in nucleic acid metabolism,nucleobase-containing compound metabolism,regulation of cellular metabolism and other biological processes;may be involved in cell nuclear,nuclear,intracellular,intracellular organelles and other cellular functions It may be involved in cellular components such as heterocyclic compound binding,organic cyclic compound binding,ATP binding,and adenine ribonucleotide binding.KEGG enrichment analysis revealed that the host gene of circulatory expression of circulatory cells in cholangiocarcinoma was associated with cancer proteoglycan,hepatocellular carcinoma,ErbB signaling pathway,thyroid hormone signaling pathway,non-small cell lung cancer,MAPK signaling pathway,and cancer miRNA.GO enrichment analysis found that cholangiocarcinoma tissue down-regulated expression of circRNAs host genes may be involved in RNA catabolism,cell macromolecular catabolism,mRNA metabolism,macromolecular catabolism and other biological processes;may be involved in cytoplasmic,intracellular and other molecular functions.It may be involved in cellular components such as protein binding,RNA binding,heterocyclic compound binding,and ATP binding.KEGG pathway enrichment analysis revealed that cholangiocarcinoma tissues downregulated host genes expressing circRNAs and Shigella infection,endoplasmic reticulum protein processing,bacterial invasion of epithelial cells,RNA degradation,TGF-? signaling pathway,neurotrophin signaling pathway,adhesion It regulates the actin cytoskeleton,ubiquitin-mediated proteolysis,and the signaling pathway regulates the pluripotency of stem cells.Conclusion: CircRNAs are differentially expressed in cholangiocarcinoma and adjacent tissues.The biosignal analysis suggests that differential expression of circRNAs may be involved in the development of cholangiocarcinoma.Part ? hsa_circRNA₀00585 expression and functional verificationin cholangiocarcinomaOBJECTIVE: To select the hsa_circRNA₀00585 expanded sample that was up-regulated in cholangiocarcinoma for real-time quantitative reverse transcription polymerase chain reaction?qRT-PCR?verification and to target its downstream targets mir-615-5p and AMOT/ The expression level of YAP was detected by qRT-PCR.Methods: The expression levels of hsa_circRNA₀00585,mir-615-5p and AMOT/YAP were detected by qRT-PCR in 15 pairs of matched cholangiocarcinoma tissues and adjacent tissues,and the differential expression of cancer tissues and adjacent tissues was compared.The Receiver Operating Characteristic?ROC?evaluated the value of hsa_circRNA₀00585 as a diagnostic biomarker for cholangiocarcinoma.RESULTS: The expression of hsa_circRNA₀00585 was significantly up-regulated in cholangiocarcinoma tissues.Consistent with the primary screening results,mir-615-5p was down-regulated significantly,and AMOT/YAP was upregulated in cholangiocarcinoma.Based on this,we hypothesized that hsa_circRNA₀00585 as mir-615-5p Molecular sponge regulates the target protein AMOT/YAP to participate in the development of cholangiocarcinoma.ROC curve evaluation hsa_circRNA₀00585 as a biomarker for the diagnosis of cholangiocarci-noma specificity and sensitivity were?AUC = 0.693?.Conclusion: hsa_circRNA₀00585 can be used as a biomarker for cholangioca-rcinoma in cholangiocarcinoma.The hsa_circRNA₀00585/mir-615-5p/ AMOT/ YAP pathway may be involved in the development of cholangiocarcinoma and may be a potential therapeutic target for cholangiocarcinoma.