乳铁蛋白对脑胶质瘤细胞生物学行为的影响研究  

Effect of lactoferrin on biological behaviors of brain glioma cells

作　　者：王晛珏 刘刚 苏丽娅[1] 刘海波[2] Wang Xianjue;Liu Gang;Su Liya;Liu Haibo(Clinical Medical Research Center,Affiliated Hospital of Inner Mongolia Medical University,Inner Mongolia Key Laboratory of Medical Cell Biology,Hohhot 010050,China;Department of Neurosurgery,Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院临床医学研究中心,内蒙古自治区医学细胞生物学重点实验室,呼和浩特010050 [2]内蒙古医科大学附属医院神经外科,呼和浩特010050

出　　处：《中华神经医学杂志》2023年第1期51-57,共7页Chinese Journal of Neuromedicine

基　　金：内蒙古自然科学基金(2019MS08020);内蒙古卫生健康科技计划项目(202201277);内蒙古医科大学致远人才计划(ZY0120025)。

摘　　要：目的探讨乳铁蛋白对脑胶质瘤细胞生物学行为的影响及其机制。方法复苏冻存的人胶质母细胞瘤细胞株U87MG后,应用MTT法检测不同浓度(0、100、200及300μg/mL)乳铁蛋白对U87MG细胞增殖能力的影响,以筛选最适药物浓度。将U87MG细胞分为空白对照组与乳铁蛋白(200μg/mL)处理组,应用Edu染色、Transwell实验、Mito-Tracker染色、DCFH-DA染色检测2组细胞的增殖情况、迁移情况及线粒体活性、活性氧生成水平,应用免疫荧光化学染色检测2组细胞中自噬标志蛋白微管相关蛋白1A/1B轻链3B(LC3B)的免疫荧光染色强度,应用化学比色法检测2组细胞中氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA)含量,应用RT-PCR法检测2组细胞中上皮-间充质转化过程相关基因(E-钙黏蛋白、纤连蛋白、N-钙黏蛋白、波形蛋白及Snail)的表达,应用Western blotting检测2组细胞中凋亡相关蛋白(Bax、Bcl-2)的表达。结果与空白对照组比较,乳铁蛋白处理组中Edu阳性细胞比例、细胞迁移数目、Mito-Tracker阳性细胞比例明显降低,DCFH-DA阳性细胞比例明显升高,细胞质中LC3B免疫荧光染色强度明显升高,SOD含量明显降低,MDA含量明显升高,E-钙黏蛋白mRNA表达明显降低,纤连蛋白、N-钙黏蛋白、波形蛋白及Snail mRNA表达明显升高,Bax蛋白表达明显升高,Bcl-2蛋白表达明显降低,差异均有统计学意义(P<0.05)。结论乳铁蛋白可有效抑制脑胶质瘤细胞的增殖、迁移、侵袭等生物学行为,其机制与调控线粒体活性密切相关。Objective To investigate the effect and mechanism of lactoferrin(LF)on biological behaviors of brain glioma cells.Methods The effect of LF at different concentrations(0,100,200 and 300μg/mL)on the proliferative ability of human glioma cell line U87MG was analyzed by MTT assay to screen the optimal drug concentration.U87MG cells were divided into blank control group and LF(200μg/mL)treatment group.Edu staining,Transwell assay,Mito-Tracker staining and DCFH-DA staining were used to detect the cell proliferation,migration,mitochondrial activity and reactive oxygen level.Autophagy marker protein microtubule associated protein 1 light chain 3B(LC3B)expression was detected by immunofluorescent chemical staining;contents of superoxide dismutase(SOD)and malonaldehyde(MDA)were detected by colorimetric chemical sensors;expressions of genes related to epithel-to-mesenchymal transformation(E-cadherin,fibronectin,N-cadherin,vimentin and Snail)were detected by reverse transcription(RT)-PCR;expressions of apoptosis-related proteins(Bax and Bcl-2)were detected by Western blotting.Results Compared with blank control group,the LF treatment group had significantly decreased proportions of Edu positive cells and Mito-Tracker positive cells,number of cell migration and SOD content,and statistically increased proportion of DCFH-DA positive cells,MDA content and LC3B immunofluorescent staining intensity in cytoplasm(P<0.05).Compared with blank control group,the LF treatment group had significantly decreased E-cadherin mRNA expression and Bcl-2 protein expression,and statistically increased fibronectin,N-cadherin,vimentin and Snail mRNA expressions and Bax protein expression(P<0.05).Conclusion LF can effectively inhibit the proliferation,migration,invasion and other biological behaviors of glioma cells,whose mechanism is closely related to mitochondrial activity.

关 键 词：神经胶质瘤 乳铁蛋白 线粒体活性 细胞实验 

分 类 号：R739.41[医药卫生—肿瘤]