miR-132靶向调控PTEN基因对雨蛙素诱导的急性胰腺炎模型胰腺腺泡细胞增殖及凋亡的影响  

Effect of mi R-132 targeting PTEN gene on proliferation and apoptosis of acute pancreatic acinar cells in a model of cerulein-induced acute pancreatitis

作　　者：王伟[1] 朱婷婷 刘家豪 WANG Wei;ZHU Tingting;LIU Jiahao(Department of Gastroenterology,Qingdao Eighth People’s Hospital,Qingdao 266000,China;Department of Minimally Invasive Interventional Therapy Center,Qingdao Municipal Hospital(East Hospital),Qingdao 266000,China)

机构地区：[1]青岛市第八人民医院消化内科,266000 [2]青岛市市立医院(东院)微创介入治疗中心,266000

出　　处：《国际消化病杂志》2022年第4期261-266,共6页International Journal of Digestive Diseases

摘　　要：目的 探究miR-132靶向调控磷酸酶与张力蛋白同源物(PTEN)基因对雨蛙素诱导的急性胰腺炎(AP)模型胰腺腺泡细胞增殖、凋亡的影响。方法 体外诱导并培养AP MZ-M0321细胞,采用ELISA法检测空白对照组和雨蛙素组(经雨蛙素处理)细胞的IL-6、TNF-α表达水平。取经雨蛙素处理的细胞随机分为无义序列组(转染miR-132-mimics-NC序列)、miR-132过表达组(转染miR-132-mimics序列)和共转染组(转染质粒载体pc DNA-PTEN+miR-132-mimics序列)。采用实时荧光定量PCR法检测各转染组miR-132、PTEN m RNA的表达水平。采用MTT法、Annexin V-FITC/PI双染法检测各组细胞的增殖、凋亡情况。采用蛋白质印迹法检测各转染组中PTEN、增殖和凋亡相关蛋白的表达水平。应用Target Scan数据库预测miR-132与PTEN的靶向关系,并采用双荧光素酶报告基因实验验证。结果 AP细胞模型构建成功。经成功转染细胞后,与无义序列组比较,miR-132过表达组细胞的增殖抑制率、凋亡率、PTEN、Bax蛋白表达水平降低,Ki-67、Bcl-2蛋白表达水平升高,差异均有统计学意义(P均<0.05);与miR-132过表达组比较,共转染组细胞的增殖抑制率、凋亡率、PTEN、Bax蛋白表达水平升高,Ki-67、Bcl-2蛋白表达水平降低,差异均有统计学意义(P均<0.05)。Target Scan数据库预测结果显示,PTEN是miR-132的潜在靶基因,双荧光素酶报告基因实验结果证实两者存在靶向关系。结论 miR-132过表达可靶向下调PTEN表达,促进雨蛙素诱导的AP MZ-M0321细胞增殖并抑制细胞凋亡。Objective This paper intends to investigate the effect of miR-132 on the proliferation and apoptosis of pancreatic acinar cells in a model of cerulein-induced acute pancreatitis by targeting PTEN gene.Methods AP MZ-M0321 cells were inducted and cultured in vitro,and the expression of IL-6 and TNF-α in the blank control group and the cerulein group(treated with cerulein) were detected by ELISA.The cerulein-treated cells were randomly assigned to the nonsense sequence group(transfected with miR-132-mimics-NC sequence),the miR-132 overexpressed group(transfected with miR-132-mimics sequence),and the co-transfection group(transfection with plasmid vector pc DNA-PTEN + miR-132-mimics sequence).The expression of miR-132 and PTEN m RNA were detected by real-time quantitative PCR.Cell proliferation and apoptosis were detected by MTT assay and Annexin V-FITC/PI double-staining.The expression of PTEN,proliferation,and apoptosis-related protein in transfected cells were detected by Western blotting.The targeting relationship between miR-132 and PTEN was predicted by Target Scan database and verified by the double luciferase reporter gene experiment.Results The AP cell model is established successfully.After successful transfection,compared with the nonsense sequence group,the proliferation inhibition rate,the apoptosis rate,the expression of PTEN and Bax protein in the miR-132 overexpression group are decreased,and the expression of Ki-67 and Bcl-2 protein are increased,with a statistically significant difference(P<0.05).Compared with the miR-132 overexpression group,the proliferation inhibition rate,the apoptosis rate,the expression of PTEN,and Bax protein in co-transfection group are increased,and the expression of Ki-67 and Bcl-2 proteins are decreased,with a statistically significant difference(P<0.05).The Target Scan database indicates that PTEN is a potential target gene for miR-132,and the double luciferase reporter gene experiment has confirmed the targeting relationship between the two.Conclusion Overexpression of

关 键 词：急性胰腺炎 细胞增殖 细胞凋亡 

分 类 号：R576[医药卫生—消化系统]