目的探讨环状RNA(circ)_0003028对人肝癌细胞增殖、迁移和侵袭的影响及分子机制。方法人肝癌细胞系Huh7分为小干扰RNA(si)-NC组、si-circ_0003028组、微小RNA(miR)-NC组、miR-498模似物(mimics)组、si-circ_0003028+anti-miR-NC组、si-c...目的探讨环状RNA(circ)_0003028对人肝癌细胞增殖、迁移和侵袭的影响及分子机制。方法人肝癌细胞系Huh7分为小干扰RNA(si)-NC组、si-circ_0003028组、微小RNA(miR)-NC组、miR-498模似物(mimics)组、si-circ_0003028+anti-miR-NC组、si-circ_0003028+anti-miR-498组;Real-time PCR检测肝癌组织及各组细胞中circ_0003028和miR-498表达水平;MTT检测细胞增殖;Transwell检测细胞迁移和侵袭数;Western blotting检测蛋白表达;双荧光素酶报告实验检测circ_0003028和miR-498的靶向调控关系。结果肝癌组织中circ_0003028表达水平升高(0.98±0.02 vs 1.36±0.01),miR-498表达水平降低(0.98±0.02 vs 0.63±0.02)(P<0.05)。抑制circ_0003028表达或miR-498过表达后,Huh7细胞中Ki-67(0.85±0.02 vs 0.41±0.02或0.95±0.11 vs 0.37±0.02)、基质金属蛋白酶(MMP)-2(0.71±0.02 vs 0.43±0.03或0.83±0.02 vs 0.41±0.03)、MMP-9(0.74±0.02 vs 0.37±0.02或0.78±0.02 vs 0.39±0.02)蛋白表达水平降低,细胞活性(1.53±0.03 vs 1.05±0.02或1.68±0.02 vs 1.11±0.02)降低;迁移(111.40±2.12 vs 77.22±2.38或108.90±2.30 vs 78.44±1.46)和侵袭(87.89±2.18 vs 49.78±1.98或80.22±1.79 vs 38.22±1.52)细胞数减少,生殖器形成抑制基因-1(SMG-1)(0.76±0.02 vs 1.39±0.02或0.79±0.02 vs 1.39±0.02)、p53(0.77±0.02 vs 1.24±0.03或0.82±0.03 vs 1.45±0.03)、p53-ser15(0.78±0.03 vs 1.50±0.02或0.82±0.02 vs 1.49±0.04)蛋白表达水平升高(P<0.05)。Circ_0003028靶向调控miR-498,沉默miR-498逆转了抑制circ_0003028表达对Huh7细胞增殖、迁移和侵袭的影响。结论抑制circ_0003028表达通过靶向miR-498影响SMG-1/p53信号通路而抑制人肝癌细胞增殖、迁移和侵袭。展开更多
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
文摘目的探讨环状RNA(circ)_0003028对人肝癌细胞增殖、迁移和侵袭的影响及分子机制。方法人肝癌细胞系Huh7分为小干扰RNA(si)-NC组、si-circ_0003028组、微小RNA(miR)-NC组、miR-498模似物(mimics)组、si-circ_0003028+anti-miR-NC组、si-circ_0003028+anti-miR-498组;Real-time PCR检测肝癌组织及各组细胞中circ_0003028和miR-498表达水平;MTT检测细胞增殖;Transwell检测细胞迁移和侵袭数;Western blotting检测蛋白表达;双荧光素酶报告实验检测circ_0003028和miR-498的靶向调控关系。结果肝癌组织中circ_0003028表达水平升高(0.98±0.02 vs 1.36±0.01),miR-498表达水平降低(0.98±0.02 vs 0.63±0.02)(P<0.05)。抑制circ_0003028表达或miR-498过表达后,Huh7细胞中Ki-67(0.85±0.02 vs 0.41±0.02或0.95±0.11 vs 0.37±0.02)、基质金属蛋白酶(MMP)-2(0.71±0.02 vs 0.43±0.03或0.83±0.02 vs 0.41±0.03)、MMP-9(0.74±0.02 vs 0.37±0.02或0.78±0.02 vs 0.39±0.02)蛋白表达水平降低,细胞活性(1.53±0.03 vs 1.05±0.02或1.68±0.02 vs 1.11±0.02)降低;迁移(111.40±2.12 vs 77.22±2.38或108.90±2.30 vs 78.44±1.46)和侵袭(87.89±2.18 vs 49.78±1.98或80.22±1.79 vs 38.22±1.52)细胞数减少,生殖器形成抑制基因-1(SMG-1)(0.76±0.02 vs 1.39±0.02或0.79±0.02 vs 1.39±0.02)、p53(0.77±0.02 vs 1.24±0.03或0.82±0.03 vs 1.45±0.03)、p53-ser15(0.78±0.03 vs 1.50±0.02或0.82±0.02 vs 1.49±0.04)蛋白表达水平升高(P<0.05)。Circ_0003028靶向调控miR-498,沉默miR-498逆转了抑制circ_0003028表达对Huh7细胞增殖、迁移和侵袭的影响。结论抑制circ_0003028表达通过靶向miR-498影响SMG-1/p53信号通路而抑制人肝癌细胞增殖、迁移和侵袭。
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.
