BACKGROUND Increasing evidence demonstrates that by acting as microRNA sponges modulating gene expression at the transcriptional or post-transcriptional level,circular RNAs(circRNAs)participate in the pathogenesis of ...BACKGROUND Increasing evidence demonstrates that by acting as microRNA sponges modulating gene expression at the transcriptional or post-transcriptional level,circular RNAs(circRNAs)participate in the pathogenesis of a variety of diseases and are considered ideal biomarkers of human disease.AIM To examine the expression of circRNA_103516 in inflammatory bowel disease(IBD)and its associations with clinical phenotypes and inflammatory cytokines.METHODS Peripheral blood mononuclear cells(PBMCs)were obtained from patients with IBD,healthy controls(HCs),and patient controls(PCs).Expression of circRNA_103516 and hsa-miR-19b-1-5p was assessed by quantitative reverse transcription-polymerase chain reaction.Crohn's disease activity index(CDAI),Mayo score,C-reactive protein(CRP)level,and erythrocyte sedimentation rate(ESR)were measured.To assess the inflammatory cytokines tumour necrosis factorα(TNF-α),interferon-γ(IFN-γ),and interleukin-10(IL-10),blood samples were analysed by flow cytometry.RESULTS Ninety Crohn’s disease(CD)and 90 ulcerative colitis(UC)patients,80 HCs,and 35 PCs were included in the study.CircRNA_103516 was upregulated in CD and UC patients compared with HCs and PCs(P<0.05).The area under the curve of circRNA_103516 for diagnosing CD and UC was 0.790 and 0.687,respectively.In addition,circRNA_103516 levels were increased in active CD and UC compared with remittent groups(P=0.027,P=0.045).Furthermore,in CD,circRNA_103516 correlated positively with CDAI(P<0.001),CRP(P<0.001),ESR(P<0.001),TNFα(P<0.001),and IFN-γ(P<0.001)and negatively correlated with IL-10(P=0.006).In UC patients,circRNA_103516 correlated with Mayo score(P<0.001),CRP(P<0.001),ESR(P<0.001),TNFα(P<0.001),IFN-γ(P=0.011),and IL-10(P=0.002).Additionally,circRNA_103516 correlated positively with stricturing(P=0.018)and penetrating(P=0.031)behaviour.Moreover,hsamiR-19b-1-5p correlated negatively with circRNA_103516 in CD.CONCLUSION CircRNA_103516 levels in PBMCs can be considered an ideal candidate biomarker for diagnosing IBD.Dysregulation of circRNA_103516 may participate in the molecular mechanism of IBD through hsa-miR-19b-1-5p sponging.展开更多
BACKGROUND The incidence of inflammatory bowel disease,a chronic intestinal inflammatory disorder that includes Crohn’s disease(CD)and ulcerative colitis,is rising.Circular RNAs are considered valuable diagnostic bio...BACKGROUND The incidence of inflammatory bowel disease,a chronic intestinal inflammatory disorder that includes Crohn’s disease(CD)and ulcerative colitis,is rising.Circular RNAs are considered valuable diagnostic biomarkers for CD.Current evidence supports the views that epithelial-mesenchymal transition(EMT)plays an important role in CD pathogenesis,and that hsa-miR-130a-3p can inhibit transforming growth factor-β1(TGF-β1)-induced EMT.Our previous study revealed that hsa_circRNA_102610 was upregulated in CD patients.Moreover,we predicted an interaction between hsa_circRNA_102610 and hsa-miR-130a-3p.Thus,we hypothesized that hsa_circRNA_102610 may play roles in the proliferation and EMT of intestinal epithelial cells by sponging hsa-miR-130a-3p to participate in the pathogenesis of CD.AIM To explore the mechanism of hsa_circRNA_102610 in the pathogenesis of CD.METHODS The relative expression levels of hsa_circRNA_102610 and hsa-miR-130a-3p in patients were detected by quantitative reverse transcription-polymerase chain reaction.The proliferation of human intestinal epithelial cells(HIECs)and normal-derived colon mucosa cell line 460(NCM460)cells was detected by cell counting kit-8,5-ethynyl-2’-deoxyuridine staining and cell cycle assays following overexpression or downregulation of hsa_circRNA_102610.Cell proliferation assays were performed as described above in a rescue experiment with hsa-miR-130a-3p mimics.The interaction of hsa_circRNA_102610 and hsa-miR-130a-3p was verified by fluorescence in situ hybridization and dual luciferase reporter assays.The relative expression levels of CyclinD1,mothers against decapentaplegic homolog 4(SMAD4),E-cadherin,N-cadherin and Vimentin were detected by western blotting following hsa_circRNA_102610 overexpression,TGF-β1-induced EMT or hsa-miR-130a-3p mimic transfection(in rescue experiments).RESULTS Upregulation of hsa_circRNA_102610 was determined to be positively correlated with elevated fecal calprotectin levels in CD(r=0.359,P=0.007)by Pearson correlation analysis.Hsa_circRNA_102610 promoted the proliferation of HIECs and NCM460 cells,while hsa-miR-130a-3p reversed the cell proliferationpromoting effects of hsa_circRNA_102610.Fluorescence in situ hybridization and dual luciferase reporter assays showed that hsa_circRNA_102610 directly bound hsa-miR-130a-3p in NCM460 and 293T cells.An inverse correlation between downregulation of hsa-miR-130a-3p and upregulation of hsa_circRNA_102610 in CD patients was observed(r=-0.290,P=0.024)by Pearson correlation analysis.Moreover,overexpression of hsa_circRNA_102610 promoted SMAD4 and CyclinD1 protein expression validated by western-blotting.Furthermore,overexpression of hsa_circRNA_102610 promoted TGF-β1 induced EMT in HIECs and NCM460 cells via targeting of hsa-miR-130a-3p,with increased expression of Vimentin and N-cadherin and decreased expression of E-cadherin.CONCLUSION Hsa_circRNA_102610 upregulation in CD patients could promote the proliferation and EMT of intestinal epithelial cells via sponging of hsa-miR-130a-3p.展开更多
基金Supported by the Natural Science Foundation of Jiangsu Province,No.BK20161232the Suzhou Special Project of Diagnosis and Treatment for Key Clinical Disease,No.LCZX201715
文摘BACKGROUND Increasing evidence demonstrates that by acting as microRNA sponges modulating gene expression at the transcriptional or post-transcriptional level,circular RNAs(circRNAs)participate in the pathogenesis of a variety of diseases and are considered ideal biomarkers of human disease.AIM To examine the expression of circRNA_103516 in inflammatory bowel disease(IBD)and its associations with clinical phenotypes and inflammatory cytokines.METHODS Peripheral blood mononuclear cells(PBMCs)were obtained from patients with IBD,healthy controls(HCs),and patient controls(PCs).Expression of circRNA_103516 and hsa-miR-19b-1-5p was assessed by quantitative reverse transcription-polymerase chain reaction.Crohn's disease activity index(CDAI),Mayo score,C-reactive protein(CRP)level,and erythrocyte sedimentation rate(ESR)were measured.To assess the inflammatory cytokines tumour necrosis factorα(TNF-α),interferon-γ(IFN-γ),and interleukin-10(IL-10),blood samples were analysed by flow cytometry.RESULTS Ninety Crohn’s disease(CD)and 90 ulcerative colitis(UC)patients,80 HCs,and 35 PCs were included in the study.CircRNA_103516 was upregulated in CD and UC patients compared with HCs and PCs(P<0.05).The area under the curve of circRNA_103516 for diagnosing CD and UC was 0.790 and 0.687,respectively.In addition,circRNA_103516 levels were increased in active CD and UC compared with remittent groups(P=0.027,P=0.045).Furthermore,in CD,circRNA_103516 correlated positively with CDAI(P<0.001),CRP(P<0.001),ESR(P<0.001),TNFα(P<0.001),and IFN-γ(P<0.001)and negatively correlated with IL-10(P=0.006).In UC patients,circRNA_103516 correlated with Mayo score(P<0.001),CRP(P<0.001),ESR(P<0.001),TNFα(P<0.001),IFN-γ(P=0.011),and IL-10(P=0.002).Additionally,circRNA_103516 correlated positively with stricturing(P=0.018)and penetrating(P=0.031)behaviour.Moreover,hsamiR-19b-1-5p correlated negatively with circRNA_103516 in CD.CONCLUSION CircRNA_103516 levels in PBMCs can be considered an ideal candidate biomarker for diagnosing IBD.Dysregulation of circRNA_103516 may participate in the molecular mechanism of IBD through hsa-miR-19b-1-5p sponging.
基金Supported by the Suzhou Special Project of Diagnosis and Treatment for Key Clinical Disease,No.LCZX201715the Natural Science Foundation of Jiangsu Province,No.BK20161232the Science and Technology Development Fund of Nanjing Medical University,No.NMUB2018215.
文摘BACKGROUND The incidence of inflammatory bowel disease,a chronic intestinal inflammatory disorder that includes Crohn’s disease(CD)and ulcerative colitis,is rising.Circular RNAs are considered valuable diagnostic biomarkers for CD.Current evidence supports the views that epithelial-mesenchymal transition(EMT)plays an important role in CD pathogenesis,and that hsa-miR-130a-3p can inhibit transforming growth factor-β1(TGF-β1)-induced EMT.Our previous study revealed that hsa_circRNA_102610 was upregulated in CD patients.Moreover,we predicted an interaction between hsa_circRNA_102610 and hsa-miR-130a-3p.Thus,we hypothesized that hsa_circRNA_102610 may play roles in the proliferation and EMT of intestinal epithelial cells by sponging hsa-miR-130a-3p to participate in the pathogenesis of CD.AIM To explore the mechanism of hsa_circRNA_102610 in the pathogenesis of CD.METHODS The relative expression levels of hsa_circRNA_102610 and hsa-miR-130a-3p in patients were detected by quantitative reverse transcription-polymerase chain reaction.The proliferation of human intestinal epithelial cells(HIECs)and normal-derived colon mucosa cell line 460(NCM460)cells was detected by cell counting kit-8,5-ethynyl-2’-deoxyuridine staining and cell cycle assays following overexpression or downregulation of hsa_circRNA_102610.Cell proliferation assays were performed as described above in a rescue experiment with hsa-miR-130a-3p mimics.The interaction of hsa_circRNA_102610 and hsa-miR-130a-3p was verified by fluorescence in situ hybridization and dual luciferase reporter assays.The relative expression levels of CyclinD1,mothers against decapentaplegic homolog 4(SMAD4),E-cadherin,N-cadherin and Vimentin were detected by western blotting following hsa_circRNA_102610 overexpression,TGF-β1-induced EMT or hsa-miR-130a-3p mimic transfection(in rescue experiments).RESULTS Upregulation of hsa_circRNA_102610 was determined to be positively correlated with elevated fecal calprotectin levels in CD(r=0.359,P=0.007)by Pearson correlation analysis.Hsa_circRNA_102610 promoted the proliferation of HIECs and NCM460 cells,while hsa-miR-130a-3p reversed the cell proliferationpromoting effects of hsa_circRNA_102610.Fluorescence in situ hybridization and dual luciferase reporter assays showed that hsa_circRNA_102610 directly bound hsa-miR-130a-3p in NCM460 and 293T cells.An inverse correlation between downregulation of hsa-miR-130a-3p and upregulation of hsa_circRNA_102610 in CD patients was observed(r=-0.290,P=0.024)by Pearson correlation analysis.Moreover,overexpression of hsa_circRNA_102610 promoted SMAD4 and CyclinD1 protein expression validated by western-blotting.Furthermore,overexpression of hsa_circRNA_102610 promoted TGF-β1 induced EMT in HIECs and NCM460 cells via targeting of hsa-miR-130a-3p,with increased expression of Vimentin and N-cadherin and decreased expression of E-cadherin.CONCLUSION Hsa_circRNA_102610 upregulation in CD patients could promote the proliferation and EMT of intestinal epithelial cells via sponging of hsa-miR-130a-3p.
