miR-451a对人胰腺癌BxPc3细胞增殖和凋亡的影响及其分子机制研究  

Effects of miR-451a on proliferation and apoptosis of human pancreatic cancer BxPc3 cells and its molecular mechanisms

作　　者：刘少朋 刘海潮 白明辉 LIU Shaopeng;LIU Haichao;BAI Minghui(The Second Department of Hepatobiliary,Pancreatic,Splenic and Hernia Surgery,Luoyang Central Hospital Affiliated to Zhengzhou University,Luoyang,Henan 471000,P.R.China)

机构地区：[1]郑州大学附属洛阳中心医院肝胆胰脾及疝外科二病区,河南洛阳471000

出　　处：《中国普外基础与临床杂志》2020年第10期1231-1235,共5页Chinese Journal of Bases and Clinics In General Surgery

基　　金：2018年市级科技医疗卫生项目(编号:1820003A)。

摘　　要：目的分析miR-451a对人胰腺癌BxPc3细胞增殖及凋亡的影响并探究其分子机制。方法以BxPc3细胞为研究对象,建立脂质体转染miR-451a模拟物并分别设不同浓度(25、50、100和200μmol/L)miR-451a组及空白对照组。实时荧光定量PCR法检测各组细胞内miR-451a mRNA表达情况;然后用MTT法、平板克隆实验、流式细胞术及Western blot法分别检测不同浓度miR-451a转染对BxPc3细胞的增殖能力、细胞克隆数、细胞周期及凋亡变化的影响以及BxPc3细胞中巨噬细胞移动抑制因子(MIF)、钙结合蛋白39(CAB39)、磷酸化磷脂酰肌醇-3激酶(p-PI3K)和磷酸化蛋白激酶B(p-AKT)蛋白的表达情况。结果各浓度转染组BxPc3细胞中miR-451a mRNA表达量高于空白对照组(P<0.050);转染miR-451a可显著抑制BxPc3细胞增殖且呈时间及浓度依赖性(P<0.050);200μmol/L miR-451a转染组细胞克隆数明显少于空白对照组(P<0.050);miR-451a转染后可使BxPc3细胞分化停滞于G0/G1期并可诱导细胞凋亡且具有浓度依赖性(P<0.050);miR-451a转染后BxPc3细胞内MIF、CAB39、p-PI3K和p-AKT蛋白表达较空白对照组下调并呈现浓度依赖性(P<0.050)。结论从本研究实验结果看,miR-451a可抑制BxPc3细胞增殖、诱导细胞凋亡并呈现浓度依赖性,其机制可能与抑制PI3K/AKT信号通路有关。Objective To analyze the effects of miR-451a on the proliferation and apoptosis of human pancreatic cancer BxPc3 cells,and to explore its molecular mechanisms.Methods The liposome transfection mimics of miR-451a were established in the BxPc3 cells,which were used as the research objects,and different concentrations(25,50,100 and 200μmol/L)of miR-451a and blank control group were set up respectively.The expression of miR-451a mRNA in the BxPc3 cells after the transfection was detected by the qRT-PCR method.The effects of miR-451a at different concentrations on the proliferation,cell clone number,cell cycle and apoptosis,and the expressions of the macrophage migration inhibitory factor(MIF),calcium binding protein 39(CAB39),phosphorylated phosphatidylinositol-3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)proteins in the BxPc3 cells were detected by the MTT assay,plate cloning assay,flow cytometry,and Western blot,respectively.Results The expressions of miR-451a mRNA in the transfected BxPc3 cells were significantly higher than in the blank control BxPc3 cells(P<0.050).The miR-451a could inhibit the proliferation of BxPc3 cells in a time-and concentration-dependent manner significantly(P<0.050),block the differentiation of BxPc3 cells in the G0/G1 phase,and induce the apoptosis with a concentration-dependent manner(P<0.050).The expressions of MIF,CAB39,p-PI3K,and p-AKT proteins in the BxPc3 cells were down-regulated with a concentration-dependent manner(P<0.050).Conclusion From results of this study,miR-451a could inhibit proliferation and induce apoptosis of BxPc3 cells in a concentration-dependent manner,and its mechanisms might be related to inhibition of PI3K/AKT signaling pathway.

关 键 词：胰腺癌 BxPc3细胞 miR-451a 磷脂酰肌醇-3激酶/磷酸化蛋白激酶B信号通路 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]