stomatin对非小细胞肺癌细胞中葡萄糖转运蛋白1表达及定位的影响  

Effect of stomatin on the expression and translocation of glucose transporter 1 in non-small cell lung cancer

作　　者：刘明依 安华英 袁伟 马洁 LIU Mingyi;AN Huaying;YUAN Wei;MAJie(State Key Laboratory of Molecular Oncology, National Cancer Center / National Clinical Research Center for Cancer / Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China;Department of Biotherapy, Beijing Hospital, National Center of Gerontology, Beijing 100730, China)

机构地区：[1]国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院分子肿瘤学国家重点实验室,北京100021 [2]北京医院生物治疗中心/国家老年医学中心,北京100730

出　　处：《癌症进展》2019年第4期399-402,共4页Oncology Progress

基　　金：国家自然科学基金(81602576)

摘　　要：目的探究stomatin基因对非小细胞肺癌细胞中葡萄糖转运蛋白1(Glut1)表达及定位的影响。方法采用RNA干扰技术敲降A549、H520细胞内stomatin基因,利用实时荧光定量聚合酶链反应技术及蛋白质免疫印迹技术检测Glut1在两种细胞内的表达变化。采用细胞免疫荧光染色技术对stomatin蛋白及Glut1蛋白进行定位并观察其位置变化。以PcDNA3.1(-)为载体,构建在Glut1第一个细胞外段插入flag标签的质粒3×flag-Glut1-PcDNA3.1(-),并将其导入过表达stomatin的A549、H520细胞中,利用流式细胞术检测细胞膜上Glut1的表达情况。结果 A549、H520细胞中敲降stomatin表达,Glut1的mRNA和蛋白表达量均未发生变化;A549、H520细胞中敲降stomatin表达,Glut1蛋白的主要表达位置由细胞膜转移到细胞质中;stomatin过表达的肺癌细胞A549、H520中,Glut1蛋白在细胞膜上的表达量明显上调。结论 stomatin表达的改变不影响非小细胞肺癌中Glut1的表达量,stomatin表达上调时能够促进Glut1在细胞膜上的定位,stomatin表达下调时Glut1更多定位在细胞质中。Objective To investigate the effect of stomatin gene on the expression and translocation of glucose transporter 1 (Glut1) in non-small cell lung cancer. Method RNA interference was used to knock down the stomatin gene in A549 and H520 cells. Then the expression of Glut1 in these cells were detected by quantitative real time polymerase chain reaction and Western blot. Cellular immunofluorescence technique was utilized to localize the stomatin protein and Glut1 protein, and to observe the translocation. The plasmid 3×flag- Glut1-PcDNA3.1(-) was constructed, which contained flag tag in the first exocellular loop of Glut1, and was transfected into A549 and H520 cells whose stomatin was overexpressed. Lastly, flow cytometry was applied to detect the expression of Glut1 which was on the cell membrane. Result Knockdown of stomatin expression in A549 and H520 cells did not affect the expression of Glut1. However, the main expression of Glut1 protein was transferred from the cell membrane to the cytoplasm. When stomatin in A549 and H520 cells were overexpressed, the expression of Glut1 protein was markedly up- regulated on cell membrane. Conclusion The expression of stomatin does not affect the expression of Glut1 in non-small cell lung cancer. The up-regulation of stomatin can promote the localization of Glut1 on the cell membrane. Glut1 is more expressed in the cytoplasm when the expression of stomatin is down-regulated.

关 键 词：非小细胞肺癌 STOMATIN 葡萄糖转运蛋白1 蛋白转位 

分 类 号：R734.2[医药卫生—肿瘤]