miR-524-5p靶向FABP5调控非小细胞肺癌增殖、凋亡的机制研究  

Role of miR-524-5p on cell proliferation and apoptosis in non-small cell lung cancer cells by targeting FABP5

作　　者：胡智慧[1] 李建芹[2] 韩鲁军[1] 段亚男[1] 张红欣[1] HU Zhi-hui;LI Jian-qin;HAN Lu-jun;DUAN Ya-nan;ZHANG Hong-xin(Department of Oncology,the First Hospital of Shijiazhuang,Shijiazhuang,Hebei 050000,China;EMG Room,Hebei Huanghua People's Hospital,Huanghua,Hebei 061100,China)

机构地区：[1]石家庄市第一医院肿瘤科,河北石家庄050000 [2]黄骅市人民医院肌电图室,河北黄骅061100

出　　处：《临床肺科杂志》2019年第11期2035-2041,共7页Journal of Clinical Pulmonary Medicine

基　　金：石家庄市卫生计划项目(No 151460703)

摘　　要：目的探讨非编码小RNA-524-5p(miR-524-5p)是否通过靶向调控脂肪酸结合蛋白5(FABP5)从而影响非小细胞肺癌的增殖和凋亡。方法实时定量PCR(qPCR)检测非小细胞肺癌细胞(A549、H1299、H1650)和正常肺上皮细胞(BEAS-2B)中miR-524-5p和FABP5表达,选择miR-524-5p表达量最低的细胞系用于后续实验;运用生物学信息预测miR-524-5p靶基因,双荧光素酶报告基因进行验证;蛋白质印迹法(Western Blot)检测上调或下调miR-524-5p对FABP5的影响,以及A549细胞中细胞周期蛋白D1(CyclinD1)、p21、B细胞淋巴瘤/白血病-2(Bcl-2)和Bax蛋白水平;采用噻唑蓝(MTT)比色法检测细胞增殖,流式细胞术检测凋亡。结果与正常肺上皮细胞相比,miR-524-5p在非小细胞肺癌细胞系中表达下调(P<0.05),FABP5 mRNA和蛋白表达上调(P<0.05)。miR-524-5p在A549细胞中的表达量最低。FABP5是miR-524-5p的靶基因。miR-524-5p过表达可抑制细胞增殖,促进细胞凋亡,降低CyclinD1、Bcl-2蛋白水平,提升p21、Bax蛋白水平,差异均达到显著水平(P<0.05),与抑制FABP5表达(si-FABP5)作用结果相同。FABP5过表达可以逆转miR-524-5p过表达对细胞增殖的抑制作用,以及对细胞凋亡的促进作用。结论miR-524-5p能够抑制非小细胞肺癌增殖,诱导细胞凋亡,其作用机制与靶向调控FABP5表达有关。Objective To explore the effect of miR-524-5p on cell proliferation and apoptosis through targeting FABP5 in non-small cell lung cancer(NSCLC).Methods NSCLC cells(A549,H1299,H1650)and normal lung epithelial cells(BEAS-2B)were collected.The miR-524-5p and FABP5 mRNA expression levels were detected by qPCR.Cell lines with the lowest expression of mir-524-5p were selected for subsequent experiments.The biological information was applied to predict the target gene of miR-524-5p,and the dual luciferase reporter gene assay was used for verification.Western blot was used to analyze the expression levels of up-regulation or down-regulation of miR-524-5p on FABP5,and expression levels of CyclinD1,p21,Bcl-2 and Bax protein.MTT assay was employed to investigate cell proliferation,and flow cytometry was adopted to testify cell apoptosis.Results The expression level of miR-524-5p was lower while FABP5 mRNA and protein were higher in NSCLC cell lines than in normal lung epithelial cells(P<0.05).The expression level of miR-524-5p in A549 cells was the lowest.FABP5 was a functional target protein of miR-524-5p.Besides,over expression of miR-524-5p,the same as inhibiting expression of FABP5(si-FABP5),significantly suppressed cell proliferation,promoted cell apoptosis,reduced the levels of CyclinD1 and Bcl-2 proteins,and increased the expression of p21 and Bax proteins(P<0.05).In addition,over expression of FABP5 reversed the influences of miR-524-5p on cell proliferation and apoptosis.Conclusion miR-524-5p inhibits cell proliferation and promotes cell apoptosis in non-small cell lung cancer by targeting and regulating FABP5.

关 键 词：非小细胞肺癌 增殖 凋亡 miR-524-5p FABP5 

分 类 号：R73[医药卫生—肿瘤]