Plectin plays a role in the migration and volume regulation of astrocytes: a potential biomarker of glioblastoma.

IF 9 2区 医学 Q1 CELL BIOLOGY
Maja Žugec, Borut Furlani, Maria J Castañon, Boštjan Rituper, Irmgard Fischer, Giuseppe Broggi, Rosario Caltabiano, Giuseppe M V Barbagallo, Michelino Di Rosa, Daniele Tibullo, Rosalba Parenti, Nunzio Vicario, Saša Simčič, Victorio Martin Pozo Devoto, Gorazd B Stokin, Gerhard Wiche, Jernej Jorgačevski, Robert Zorec, Maja Potokar
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引用次数: 0

Abstract

Background: The expression of aquaporin 4 (AQP4) and intermediate filament (IF) proteins is altered in malignant glioblastoma (GBM), yet the expression of the major IF-based cytolinker, plectin (PLEC), and its contribution to GBM migration and invasiveness, are unknown. Here, we assessed the contribution of plectin in affecting the distribution of plasmalemmal AQP4 aggregates, migratory properties, and regulation of cell volume in astrocytes.

Methods: In human GBM, the expression of glial fibrillary acidic protein (GFAP), AQP4 and PLEC transcripts was analyzed using publicly available datasets, and the colocalization of PLEC with AQP4 and with GFAP was determined by immunohistochemistry. We performed experiments on wild-type and plectin-deficient primary and immortalized mouse astrocytes, human astrocytes and permanent cell lines (U-251 MG and T98G) derived from a human malignant GBM. The expression of plectin isoforms in mouse astrocytes was assessed by quantitative real-time PCR. Transfection, immunolabeling and confocal microscopy were used to assess plectin-induced alterations in the distribution of the cytoskeleton, the influence of plectin and its isoforms on the abundance and size of plasmalemmal AQP4 aggregates, and the presence of plectin at the plasma membrane. The release of plectin from cells was measured by ELISA. The migration and dynamics of cell volume regulation of immortalized astrocytes were assessed by the wound-healing assay and calcein labeling, respectively.

Results: A positive correlation was found between plectin and AQP4 at the level of gene expression and protein localization in tumorous brain samples. Deficiency of plectin led to a decrease in the abundance and size of plasmalemmal AQP4 aggregates and altered distribution and bundling of the cytoskeleton. Astrocytes predominantly expressed P1c, P1e, and P1g plectin isoforms. The predominant plectin isoform associated with plasmalemmal AQP4 aggregates was P1c, which also affected the mobility of astrocytes most prominently. In the absence of plectin, the collective migration of astrocytes was impaired and the dynamics of cytoplasmic volume changes in peripheral cell regions decreased. Plectin's abundance on the plasma membrane surface and its release from cells were increased in the GBM cell lines.

Conclusions: Plectin affects cellular properties that contribute to the pathology of GBM. The observed increase in both cell surface and released plectin levels represents a potential biomarker and therapeutic target in the diagnostics and treatment of GBMs.

Plectin 在星形胶质细胞的迁移和体积调节中发挥作用:胶质母细胞瘤的潜在生物标志物。
背景:在恶性胶质母细胞瘤(GBM)中,水通道蛋白4(AQP4)和中间丝蛋白(IF)的表达发生了改变,但基于IF的主要细胞连接蛋白plectin(PLEC)的表达及其对GBM迁移和侵袭性的贡献尚不清楚。在此,我们评估了 Plectin 在影响星形胶质细胞质膜 AQP4 聚集分布、迁移特性和细胞体积调节方面的贡献:方法:利用公开数据集分析了人GBM中胶质纤维酸性蛋白(GFAP)、AQP4和PLEC转录本的表达,并通过免疫组化确定了PLEC与AQP4和GFAP的共定位。我们在野生型和 plectin 缺失型原代和永生化小鼠星形胶质细胞、人类星形胶质细胞和源自人类恶性 GBM 的永久细胞系(U-251 MG 和 T98G)上进行了实验。小鼠星形胶质细胞中 plectin 同工酶的表达通过实时定量 PCR 进行评估。转染、免疫标记和共聚焦显微镜被用来评估 plectin 诱导的细胞骨架分布的改变、plectin 及其异构体对质膜 AQP4 聚集体的丰度和大小的影响以及质膜上 plectin 的存在。用酶联免疫吸附法测定了细胞中plectin的释放。伤口愈合试验和钙黄绿素标记法分别评估了永生化星形胶质细胞的迁移和细胞体积的动态调节:结果:在肿瘤脑样本中,plectin和AQP4在基因表达和蛋白定位水平上呈正相关。缺乏 plectin 会导致质膜 AQP4 聚集体的丰度和大小下降,并改变细胞骨架的分布和捆绑。星形胶质细胞主要表达 P1c、P1e 和 P1g plectin 异构体。与浆膜 AQP4 聚集相关的主要 plectin 异构体是 P1c,它也对星形胶质细胞的流动性产生了最显著的影响。在缺乏 Plectin 的情况下,星形胶质细胞的集体迁移会受到影响,外围细胞区域的细胞质体积动态变化也会减弱。在 GBM 细胞系中,Plectin 在质膜表面的丰度及其从细胞中的释放均有所增加:结论:Plectin会影响细胞特性,从而导致GBM的病理变化。观察到的细胞表面和释放的 Plectin 水平的增加是诊断和治疗 GBM 的潜在生物标志物和治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Biomedical Science
Journal of Biomedical Science 医学-医学:研究与实验
CiteScore
18.50
自引率
0.90%
发文量
95
审稿时长
1 months
期刊介绍: The Journal of Biomedical Science is an open access, peer-reviewed journal that focuses on fundamental and molecular aspects of basic medical sciences. It emphasizes molecular studies of biomedical problems and mechanisms. The National Science and Technology Council (NSTC), Taiwan supports the journal and covers the publication costs for accepted articles. The journal aims to provide an international platform for interdisciplinary discussions and contribute to the advancement of medicine. It benefits both readers and authors by accelerating the dissemination of research information and providing maximum access to scholarly communication. All articles published in the Journal of Biomedical Science are included in various databases such as Biological Abstracts, BIOSIS, CABI, CAS, Citebase, Current contents, DOAJ, Embase, EmBiology, and Global Health, among others.
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