上调细胞周期相关蛋白-1的表达对脑胶质瘤U251细胞的增殖和迁移能力的影响  

Upregulated expression of Caprin-1 increases the proliferation and migration of glioma U251 cells

作　　者：严立冬 桂卉 杨卓顺 邹丹丹 王珏[2] 张力 黄宽明 罗杰 

机构地区：[1]湖北医药学院附属医院(十堰市太和医院)神经外科一病区,十堰医学硕士研究生442008 [2]湖北医药学院病理教研室,十堰442000

出　　处：《医学研究生学报》2018年第1期13-18,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81671831);十堰市科学技术研究与开发项目(17Y06)

摘　　要：目的细胞周期相关蛋白-1(Caprin-1)与肿瘤的发展进程及药物耐受密切相关。文中旨在探讨Caprin-1在脑胶质瘤临床标本中的表达情况及其对脑胶质瘤细胞系生物学特性的影响。方法选取湖北医药学院附属十堰市太和医院神经外科2015年12月至2017年4月手术切除的29份脑胶质瘤肿瘤组织和2份因外伤死亡尸检的正常脑组织标本。以U251细胞作为实验对象,构建Caprin-1高表达的U251细胞稳转株。实验分为3组:空白对照组(无转染的野生型U251细胞)、阴性对照组(转染p EGFP-C1质粒的U251细胞)、实验组(转染p EGFP-C1-Caprin-1质粒的U251细胞)。RT-PCR和Western blot分别检测3组细胞Caprin-1 mRNA和蛋白的表达水平,检测其增殖能力变化及划痕实验和Transwell小室迁移实验检测其迁移能力的变化。结果正常脑组织Caprin-1表达量(65.04±1.07)低于WHOⅡ、WHOⅢ、WHOⅣ胶质瘤的表达量(145.9±22.0、444.4±110.0、1661.0±54.5),WHOⅡ、WHOⅢ、WHOⅣCaprin-1表达量依次升高,差异有统计学意义(P<0.05)。实验组Caprin-1 mRNA、Caprin-1蛋白表达[(1.70±0.19)、(1.07±0.09)]明显高于空白对照组[(0.89±0.10)、(0.52±0.04)]、阴性对照组[(0.98±0.08)、(0.58±0.03)],差异有统计学意义(P<0.05)。实验组A值(2.55±0.14)明显高于空白对照组(1.40±0.06)和阴性对照组(1.35±0.04),差异有统计学意义(P<0.001)。实验组在每个视野下迁移的细胞数[(526.00±42.19)个]明显多于空白对照组和阴性对照组[(289.00±29.24)、(279.00±32.48)个],差异有统计学意义(P<0.001)。实验组中Cyclin D1和AKT磷酸化水平表达量明显高于空白对照组和阴性对照组(P<0.000)。结论 Caprin-1在胶质瘤标本中的表达量随肿瘤组织的恶性程度增加而增高,且上调U251细胞Caprin-1的表达则其增殖和迁移能力提高,为胶质瘤的基因靶向治疗及评估胶质瘤的预后提供理论依据。Objective Cell cycle-associated protein 1 （Caprin-1） is closely related to the development and progression of cancer. This study aimed to explore the expression of Caprin-1 in the clinical glioma specimen and its influence on the biological characteristics of the glioma cell line.Methods Brain tissue specimens were collected from 29 glioma patients and 2 normal humans that died of accidental trauma. A stably transfected U251 cell line with overexpressed Caprin-1 was established, and the U251 cells were transfected with the pEGFP-C1 plasmid （the negative control group）, or the pEGFP-C1-Caprin-1 plasmid （the experimental group）, or left untransfected （the blank control group）. The expressions of Caprin-1 mRNA and protein in the cells were determined by RT-PCR and Western blot, and the proliferation and migration of the cells examined by scratch test and Transwell assay, respectively.Results The expression of Caprin-1 was upregulated with the increased grade of glioma, 145.9±22.0, 444.4±110.0, and 1661.0±54.5 in WHO grade Ⅱ, Ⅲ, and Ⅳ glioma, respectively, significantly higher than in the normal brain tissue （P〈0.05）. Both the mRNA and protein expressions of Caprin-1 were remarkably higher in the experimental group （1.70±0.19 and 1.07±0.09） than in the blank control （0.89±0.10 and 0.52±0.04） and negative control （0.98±0.08 and 0.58±0.03） （P〈0.05）. The A value was also markedly higher in the former group （2.55±0.14） than in the latter two （1.40±0.06 and 1.35±0.04） （P〈0.01）, and so were the count of migrated cells （526.00±42.19 vs 289.00±29.24 and 279.00±32.48, P〈0.01） and the expression of CyclinD1 （0.60±0.05 vs 0.13±0.03 and 0.15±0.05, P〈0.01）.Conclusion The expression of Caprin-1 in the U251 cells was upregulated with the increased WHO grade of glioma, and the overexpression of Caprin-1 accelerated the proliferation and migration of the U251 cells.

关 键 词：脑胶质瘤 U251细胞 Caprin-1 增殖 迁移 

分 类 号：R739.41[医药卫生—肿瘤]