Manelli-Oliveira R, Machado-Santelli G M
Department of Histology and Embryology, Institute of Biomedical Sciences, University of São Paulo (USP), Av. Prof. Lineu Prestes, 1524, São Paulo, SP, CEP 05508-900, Brazil.
Histochem Cell Biol. 2001 May;115(5):403-11. doi: 10.1007/s004180100262.
Tumor cells generally present various types of nuclear alterations, which can be associated with genetic instability. The origin and mechanism of formation of the nuclear alterations are largely unknown, with the micronucleus being the most well studied alteration. The purpose of this study was to characterize the cytoskeleton filaments and to analyze the possible association between nuclear alterations and the cytoskeleton in the human lung carcinoma cells HK2 and A549. The cytoskeleton analysis was performed by using antibodies against lamin B, vimentin, cytokeratin-8, and alpha-tubulin and the secondary antibody labeled with FITC. The analysis of the actin filament was made with phalloidin-TRITC. The analyses of cytoskeleton were performed from optical sections obtained by confocal laser scanning microscopy. Filaments of the cytoskeleton of tumor cells present some differences in their distribution pattern and their expression when compared with the filaments of normal cells. The HK2 cells presented actin fibers arranged either concentrically or in clusters and tubulin filaments arranged radially, while in the A549 cells the distribution pattern was similar to that of normal cells. The lamin B filaments were the most important to identify nuclear alterations. These alterations in cytoskeleton distribution could not be associated with nuclear alterations.
肿瘤细胞通常呈现出各种类型的核改变,这可能与基因不稳定有关。核改变的起源和形成机制在很大程度上尚不清楚,其中微核是研究最为深入的改变。本研究的目的是表征细胞骨架丝,并分析人肺癌细胞HK2和A549中核改变与细胞骨架之间可能存在的关联。通过使用抗层粘连蛋白B、波形蛋白、细胞角蛋白-8和α-微管蛋白的抗体以及用异硫氰酸荧光素(FITC)标记的二抗来进行细胞骨架分析。用四甲基罗丹明异硫氰酸酯(TRITC)标记的鬼笔环肽对肌动蛋白丝进行分析。细胞骨架分析是从共聚焦激光扫描显微镜获得的光学切片上进行的。与正常细胞的丝相比,肿瘤细胞的细胞骨架丝在其分布模式和表达上存在一些差异。HK2细胞呈现出同心排列或成簇排列的肌动蛋白纤维以及径向排列的微管蛋白丝,而A549细胞中的分布模式与正常细胞相似。层粘连蛋白B丝对于识别核改变最为重要。细胞骨架分布的这些改变与核改变无关。