miR-490-5p调节低氧诱导的喉癌FaDu细胞炎症反应和血管生成的体外研究  

In Vitro Study of miR-490-5p Regulating Inflammation and Angiogenesis in Hypoxy-induced Laryngeal Carcinoma FaDu Cells

作　　者：谭元元[1] 艾合买提江·艾力江 周慧银 TAN Yuanyuan;Aihemaitijiang Ailijiang;ZHOU Huiyin(Ear,Nose and Throat Diagnosis Cancer,People’s Hospital of Xinjiang Uygur Autinimius Region,Urumqi,830000,China)

机构地区：[1]新疆维吾尔自治区人民医院耳鼻喉诊疗中心,乌鲁木齐市830000

出　　处：《医学分子生物学杂志》2025年第2期101-107,共7页Journal of Medical Molecular Biology

基　　金：新疆维吾尔自治区自然科学基金(No.2021D01C187)。

摘　　要：目的探究微小RNA-490-5p(miR-490-5p)对低氧诱导下人喉癌细胞炎症反应和血管生成的影响及分子机制。方法人下咽鳞状细胞癌细胞株FaDu分为对照组(常氧环境培养)、低氧组(低氧环境培养)、低氧+NC组(低氧环境培养+转染mimics NC)、低氧+miR-490-5p组(低氧环境培养+转染miR-490-5p mimics)。实时荧光定量PCR(RT-qPCR)检测miR-490-5p表达,CCK-8法检测细胞增殖活性,ELISA实验测定细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素IL-1β和IL-6水平,蛋白质印迹检测细胞中核因子κB(NF-κB)途径相关蛋白质表达,体外血管生成实验检测HUVEC管形成数目,RT-qPCR和蛋白质印迹检测细胞中血管内皮细胞生长因子(VEGF)、表皮生长因子(EGF)、缺氧诱导因子-1α(HIF-1α)的表达。结果与对照组比较,低氧组FaDu细胞中miR-490-5p相对表达量显著下调(P<0.05),增殖活性显著升高(P<0.05),上清液中TNF-α、IL-1β和IL-6含量显著增加(P<0.05),磷酸化NF-κB P65(p-NF-κB P65)和磷酸化IκB激酶α(p-IκBα)蛋白相对表达量显著上调(P<0.05),HUVEC管形成数目显著增加(P<0.05),VEGF、EGF、HIF-1α的mRNA和蛋白质水平显著上调(P<0.05);与低氧+NC组比较,低氧+miR-490-5p组FaDu细胞中miR-490-5p相对表达量显著上调(P<0.05),增殖活性显著降低(P<0.05),上清液中TNF-α、IL-1β和IL-6含量显著减少(P<0.05),p-NF-κB P65和p-IκBα相对表达量显著下调,同时,HUVEC管形成数目显著减少(P<0.05),VEGF、EGF、HIF-1α的mRNA和蛋白质水平也显著下调(P<0.05)。结论在低氧环境下,过表达miR-490-5p能够抑制喉癌细胞的炎症反应和血管生成。Objective To investigate the effect and molecular mechanism of microRNA-490-5p(miR-490-5p)on inflammatory response and angiogenesis in human laryngeal cancer cells induced by hypoxia.Methods The human hypopharyngeal squamous cell carcinoma cell line FaDu was divided into 4 groups:control group(cultured in normoxic environment),hypoxic group(cultured in hypoxic environment),hypoxic+NC group(cultured in hypoxic environment and then transfected with mimics NC),and hypoxic+miR-490-5p group(cultured in hypoxic environment and then transfected with miR-490-5p mimics).The expression level of miR-490-5p in cells was detected by RT-qPCR,the cell proliferation activity was detected by CCK-8 method,the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 in the cell supernatant were determined by ELISA,the expression levels of proteins related to the nuclear factorκB(NF-κB)pathway in cells was detected by Western blotting,the number of HUVEC tubes formed was detected by in vitro angiogenesis assay,the expression levels of vascular endothelial growth factor(VEGF),epidermal growth factor(EGF)and hypoxia-inducible factor-1α(HIF-1α)in cells was detected by RTqPCR and Western blotting.Results Compared with those in the control group,the relative expression level of miR-490-5p in the hypoxia group was down-regulated(P<0.05),the proliferative activity was increased(P<0.05),the levels of TNF-α,IL-1βand IL-6 in supernatant were increased(P<0.05),the relative expression levels of phosphorylated NF-κB P65(p-NF-κB P65)and phosphorylated IκB kinaseα(p-IκBα)were up-regulated(P<0.05),and the number of HUVEC tubes was increased(P<0.05),the mRNA and protein relative expression levels of VEGF,EGF and HIF-1αwere up-regulated(P<0.05).Compared with those in the hypoxia+NC group,the relative expression level of miR-490-5p in the hypoxia+miR-490-5p group was up-regulated(P<0.05),the proliferation activity was decreased(P<0.05),and the levels of TNF-α,IL-1βand IL-6 in supernatant were decreased(P<0.05),the relative

关 键 词：喉癌 低氧 miR-490-5p 炎症 血管生成 

分 类 号：R739.65[医药卫生—肿瘤]