Difference Analysis Of Circular RNA Expression Profiles Of Kaposi's Sarcoma

Objective:Screening for circRNA differences in tumor tissues and adjacent tissues of patients with Kaposi's sarcoma using gene chip technology,enriching and predicting the biological functions of differential circRNAs,and discussing the possible biological functions and values of differential circRNAs;To further study the regulatory mechanism and cure target of Kaposi's sarcoma in the field of non-coding RNA,and provide theoretical support.Methods:Five groups of cryopreserved Kaposi's sarcoma patients' tumor and adjacent tissue specimens?male: female=2:3?were selected.After checking the quality of the specimens,total RNA was extracted and reverse transcribed into cDNA.After hybridization using circRNA chips,further scans were screened to identify the difference.Heterosexual circRNA?screening criteria: difference multiple ?1.5 times,P<0.05?;circRNA is screened by calculating the P value,and the differential circRNA is imported into the circBase database,and circNet,circ2 Trait database to find its transcript,mother gene,source chromosome and Region,predict the miRNA binding site of differential circRNA,and construct the circRNA-miRNA-gene_symbol?source mother gene?expression network to predict the interaction between miRNAs and human protein-coding genes,long-chain non-coding genes,and circular RNA,Constructed an interaction network,and applied gene ontology?GO?analysis and KEGG enrichment analysis to the protein-coding genes in the miRNAs-circRNA interaction group to enrich differential circRNAs for possible biological functions Signal conditioning path.Five circRNAs were selected for reverse transcription real-time fluorescence quantitative polymerase chain reaction to verify the stability of circRNA differential expression and the reliability of gene chip screening results.Results:1.There were 550 circRNAs expressed in Kaposi's Sarcoma tumor tissue than those of tumor side tissue,of which 168 upwardly raised circRNAs and 382 downwardcircRNAs were concentrated in the exobiome region?screening criteria: P <0.05,and the difference multiple changed not less than 1.5 times?.2.Through GO analysis,it was shown that the differentially expressed circRNA is mainly enriched in R-SMAD protein binding,branching in salivary gland morphogenesis,positive regulation of focal adhesion components,very low density lipoprotein particles,positive regulation of cell junction assembly,salivary glands Development,cerebellar cortex formation,salivary gland morphogenesis,sarcomere tissue,basal plate.The results of KEGG enrichment analysis indicate that the circRNA with obvious differential expression is mainly enriched in the pathways related to substance metabolism such as glycosaminoglycan degradation,unsaturated fatty acid biosynthesis,circadian rhythm,vitamin digestion and absorption,and ECM-receptor interaction.3.Select 21 differential circRNAs with high concentration,the first 100 differential circRNAs,and the differences predict the potential target miRNA of differential circRNAs through the target prediction program,and relate to the network map with the source parent gene and constitute circRNA-miRNA-gene_symbol,and find miRNA with multiRNcircas common regulation: 676-5p,miRNA-5193.A differential hsa_circ₀092207 found on the Y chromosome was down-regulated.The difference is more significant hsa_circ₀020917,the combination of hsa-miR-575,hsa-miR-4676-5p,hsa-miR-636,hsa-miR-4293,hsa-miR-218-1-3p,111 previous differences Sexual studies have shown differences in dozens of human neoplastic diseases.Low expression hsa_circ₀022428 and hsa-miR-635 $ 66 hsa-miR-6774-5p $ 57 hsa-miR-4253 $ 53 hsa-miR-6862-5p $ 52 hsa-miR-3945 $ 51 and other five predicted binding miRNAs,all of which have more than 50 Binding site.4.5 representative circRNA?hsa_circ₀020917,hsa_circ₀022393,hsa_circ₀025653,hsa_circ₀040626,hsa_circ₀085087?line qRT-PCR verification were selected,the differential results were consistent with the differences measured by the chip in the original sample,the differential circRNA expression was stable,and the chip quality was reliable.Conclusions :1.In this study,it was found that there are differentially expressed circRNAs in KS tissues and adjacent tissues.After analysis,annotation,and correlation,it is speculated that the differentially expressed circRNAs may be involved in the pathogenesis of KS,suggesting that circRNA may be a brand new Kappa.Western sarcoma markers and potential therapeutic targets.2.Differentially expressed circRNAs are mainly enriched in molecules that regulate cell differentiation and maturation and metabolic pathways.3.The down-regulation of hsa_circ₀092207 on Y chromosome may be related to the incidence of male patients.4.hsa_circ₀020917 may be a potential biomarker molecule.