Cheng Chiung-Chi, Lai Yen-Chang Clark, Lai Yih-Shyong, Chao Wei-Ting, Tseng Yu-Hui, Hsu Yung-Hsiang, Chen You-Yin, Liu Yi-Hsiang
Department of Pathology, Chang Bing Show Chwan Memorial Hospital, Lukang, Taiwan, R.O.C. Center for General Education, Providence University, Taichung City, Taiwan, R.O.C.
Department of Pathology, Kaohsiung Medical University Hospital, Kaohsiung City, Taiwan, R.O.C.
In Vivo. 2016;30(5):549-55.
BACKGROUND/AIM: Nucleoskeleton maintains the framework of a cell nucleus that is required for a variety of nuclear functions. However, the nature of nucleoskeleton structure has not been yet clearly elucidated due to microscopy visualization limitations. Plectin, a nuclear pore-permeable component of cytoskeleton, exhibits a role of cross-linking between cytoplasmic intermediate filaments and nuclear lamins. Presumably, plectin is also a part of nucleoskeleton. Previously, we demonstrated that pleomorphism of hepatoma cells is the consequence of cytoskeletal changes mediated by plectin deficiency. In this study, we applied a variety of technologies to detect the cytoskeletons in liver cells.
The images of confocal microscopy did not show the existence of plectin, intermediate filaments, microfilaments and microtubules in hepatic nuclei. However, in the isolated nuclear preparation, immunohistochemical staining revealed positive results for plectin and cytoskeletal proteins that may contribute to the contamination derived from cytoplasmic residues. Therefore, confocal microscopy provides a simple and effective technology to observe the framework of nucleoskeleton. Accordingly, we verified that cytoskeletons are not found in hepatic cell nuclei. Furthermore, the siRNA-mediated knockdown of plectin in liver cells leads to collapsed cytoskeleton, cell transformation and pleomorphic nuclei.
Plectin and cytoskeletons were not detected in the nuclei of liver cells compared to the results of confocal microscopy. Despite the absence of nuclear plectin and cytoskeletal filaments, the evidence provided support that nuclear pleomorphism of cancer cells is correlated with the cytoplasmic disorganization of cytoskeleton.
背景/目的：核骨架维持着细胞核的框架结构，而细胞核框架是多种核功能所必需的。然而，由于显微镜可视化的局限性，核骨架结构的本质尚未得到明确阐释。网蛋白是一种可透过核孔的细胞骨架成分，在细胞质中间丝和核纤层之间发挥交联作用。据推测，网蛋白也是核骨架的一部分。此前，我们证明肝癌细胞的多形性是由网蛋白缺乏介导的细胞骨架变化所致。在本研究中，我们应用了多种技术来检测肝细胞中的细胞骨架。
共聚焦显微镜图像未显示肝细胞核中网蛋白、中间丝、微丝和微管的存在。然而，在分离的细胞核制剂中，免疫组织化学染色显示网蛋白和细胞骨架蛋白呈阳性结果，这可能是由于细胞质残留污染所致。因此，共聚焦显微镜提供了一种简单有效的技术来观察核骨架的框架结构。据此，我们证实肝细胞核中不存在细胞骨架。此外，siRNA介导的肝细胞中网蛋白敲低导致细胞骨架塌陷、细胞转化和核多形性。
与共聚焦显微镜结果相比，在肝细胞核中未检测到网蛋白和细胞骨架。尽管细胞核中网蛋白和细胞骨架丝不存在，但所提供的证据支持癌细胞的核多形性与细胞骨架的细胞质紊乱相关。
