Circular RNA Expression Profile And Preliminary Study On Function And Mechanism Of Circ0008259 In Keloid

Objective:To investigate the expression profile of circRNAs and analyze their role in keloid.Method:1.Collagenase digestion was used to isolate primary dermal fibroblasts from keloids and normal skin.2.Bioinformatic analyses and high-throughput RNA sequencing technology were applied to explore the expression profile of circRNAs in 3 human KDFs and normal dermal fibroblasts(NDFs).3.The differentially expressed circRNAs were verified by reverse transcription PCR(RT-PCR),quantitative real-time-PCR(qRT-PCR)and Sanger sequencing.4.A circRNA-microRNA(miRNA)-mRNA interaction network was created using bioinformatics tools.5.circ0008259,was selected to confirm its function by lentivirus transfection,qRT-PCR and western blot in KDFs.Results:1.Collectively,411 circRNAs,of which 206 were upregulated and 205 decreased,were found to be differentially expressed in KDFs and could bind to 2532 miRNA response elements(MREs).GO and KEGG pathways enrichment analyses showed that differentially expressed circRNAs were mainly involved in apoptosis,focal adhesion,PI3K-Akt and metabolic pathway,and may regulate the pathogenesis and development of keloid.2.Cyclization of circRNAs(circ0002198,circ0005320,circ0008259,circ0000994,circ0005480,circ0047270,circ0000375)were verified in 12 KDFs and12 NDFs using RT-PCR,Sanger sequencing.Two candidate circRNAs(circ0008259,circ0005480)were verified to be significantly reduced in KDFs,and one candidate circRNA(circ0002198)was significantly elevated in accordance with RNA-Seq dataanalysis.3.Overexpression of circ0008259 inhibited type I and ? collagen expression.Conclusion:1.The study confirmed that a large number of circRNAs were differentially expressed in keloids.Bioinformatics analysis found that differentially expressed circRNAs may be involved in the development of keloid.2.circ0008259 may play an inhibitory role in the formation of keloid collagen.