miR-7a-5p对急性胰腺炎腺泡细胞增殖、凋亡的影响及机制  

Effects of miR-7a-5p expression on proliferation and apoptosis of acinar cells in acute pancreatitis

作　　者：楼一波 王晓华[1] 傅志成 Yi-Bo Lou;Xiao-Hua Wang;Zhi-Cheng Fu(Department of Emergency Medicine,Yiwu Central Hospital,Yiwu 322000,Zhejiang Province,China;Department of Gastroenterology,Yiwu Central Hospital,Yiwu 322000,Zhejiang Province,China)

机构地区：[1]义乌市中心医院急诊科,浙江省义乌市322000 [2]义乌市中心医院消化科,浙江省义乌市322000

出　　处：《世界华人消化杂志》2019年第16期991-998,共8页World Chinese Journal of Digestology

摘　　要：背景胰腺腺泡细胞增殖及凋亡与急性胰腺炎(acute pancreatitis,AP)发生密切相关,miRNA可通过调控靶基因表达从而参与细胞增殖及凋亡过程,因而寻找与胰腺腺泡细胞增殖及凋亡相关的miRNA分子标志物对临床诊断及治疗AP具有重要意义.目的探讨微小RNA-7a-5p(miR-7a-5p)对AP腺泡细胞增殖、凋亡的影响及机制.方法构建雨蛙肽诱导的AP模型并收集胰腺炎腺泡细胞AR42J(雨蛙肽组),采用qRT-PCR与Western blot分别检测未经雨蛙肽诱导的AR42J细胞(对照组)及雨蛙肽组AR42J细胞中miR-7a-5p相对表达量及活化信号转导和转录激活因子的蛋白抑制因子-1(proteininhibitor of activated signal transducer and activator oftranscription 1,PIAS1)表达.分别将anti-miR-7a-5p、pcDNA-PIAS1转染至雨蛙肽组AR42J细胞,采用MTT法检测AR42J细胞增殖能力,流式细胞术检测AR42J细胞凋亡.荧光素酶报告系统检测miR-7a-5p对PIAS1基因的靶向调控作用,并采用Western blot检测miR-7a-5p对PIAS1蛋白表达的调控作用.采用RNA干扰技术沉默PIAS1表达(si-PIAS1组),分别将si-PIAS1及其阴性对照转染至anti-miR-7a-5p组AR42J细胞,观察AR42J细胞增殖及凋亡能力.结果与对照组相比,雨蛙肽组AR42J细胞中miR-7a-5p表达水平显著升高(P<0.05),PIAS1蛋白表达显著降低(P<0.05);抑制miR-7a-5p表达可促进胰腺炎腺泡细胞AR42J增殖并抑制其凋亡;miR-7a-5p可负向调控靶基因PIAS1表达;PIAS1过表达可促进AR42J细胞增殖并抑制其凋亡;与anti-miR-7a-5p+si-NC组相比,anti-miR-7a-5p+si-PIAS1组AR42J细胞活性显著降低(P<0.05),细胞凋亡率显著升高(P<0.05).结论miR-7a-5p可通过抑制PIAS1表达进而促进AP腺泡细胞凋亡并降低细胞增殖能力.BACKGROUND Pancreatic acinar cell proliferation and apoptosis are closely related to the development of acute pancreatitis(AP).MicroRMAs(miRNAs)participate in cell proliferation and apoptosis by regulating target gene expression,and identification of miRNA molecules related to pancreatic acinar cell proliferation and apoptosis is important for clinical diagnosis and treatment of AP.AIM To investigate the effect of miRNA-7a-5p on the proliferation and apoptosis of acinar cells in AP and the underlying mechanism.METHODS A caerulin-induced AP model was constructed using pancreatitis acinar AR42J cells.qRT-PCR and Western blot were used to detect the expression of miR-7a-5p and protein inhibitor of activated signal transducer and activator of transcription 1(PIAS1)in control AR42J cells and cerulein induced AR42J cells.After anti-miR-7a-5p and pcDNA-PIAS1 were transfected into AR42J cells,the proliferation of AR42J cells was detected by MTT assay,and the apoptosis of AR42J cells was detected by flow cytometry.The luciferase reporter system was used to detect the targeted regulation of PIAS1 gene by miR-7a-5p,and Western blot was used to detect the regulation of PIAS1 protein expression by miR-7a-5p.To silence PIAS1 expression by RNA interference,si-PIAS1 and its negative control plasmid were transfected into antimiR-7a-5p treated AR42J cells,and the proliferation and apoptosis of AR42J cells were detected.RESULTS Compared with control AR42J cells,the expression level of miR-7a-5p was significantly increased in cerulein induced AR42J cells(P<0.05),and the expression of PIAS1 protein was significantly decreased(P<0.05).Inhibition of miR-7a-5p expression promoted proliferation and inhibited apoptosis of AR42J cells.MiR-7a-5p could negatively regulate the expression of its target gene PIAS1.Overexpression of PIAS1 promoted proliferation and inhibited apoptosis of AR42J cells.Compared with the anti-miR-7a-5p+si-NC group,the activity of AR42J cells in the anti-miR-7a-5p+si-PIAS1 group was significantly decreased(P<0.05),an

关 键 词：miR-7a-5p 急性胰腺炎 胰腺腺泡细胞 增殖 凋亡 

分 类 号：R576[医药卫生—消化系统]