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目的 比较人恶性胶质瘤细胞系CHG 5和SHG 44的细胞骨架系统成分 ,以进一步了解胶质瘤细胞骨架特征与其分化程度的关系。方法 利用免疫荧光细胞化学染色结合激光共聚焦显微镜观察两种细胞系微管、微丝、中间丝的含量及分布 ,并比较几种常用固定剂和缓冲液对各骨架成分检测的影响。结果 除微管和微丝均被染色外 ,所有细胞均表达Vimentin(波形蛋白 ) ,而只有部分细胞呈胶质纤维酸性蛋白 (GFAP)阳性。微管及中间丝在两种细胞胞质中的分布特征基本相似 ,但两种细胞微丝的定位、分布及荧光强度有一定差异。Vimentin型中间丝在SHG 44细胞中表达较强 ,而GFAP在CHG 5细胞中表达较强。丙酮、75 %酒精和4%多聚甲醛对微管的固定效果较好 ;用醛类固定后中间丝 (尤其是Vimentin)染色极弱。结论 CHG 5分化程度较SHG 44高 ,两种细胞系细胞骨架状态的差异可能是二者生物学特性差异的结构基础
Objective To compare the cytoskeleton components of human malignant glioma cell lines CHG 5 and SHG 44 to further understand the relationship between cytoskeletal characteristics of glioma cells and their differentiation. Methods The content and distribution of microtubules, microfilaments and intermediate filaments in two cell lines were observed by immunofluorescence cytochemistry and laser confocal microscopy. The effects of several commonly used fixatives and buffers on the detection of skeletal components were compared. Results All cells expressed Vimentin except microtubules and microfilaments, while only some of them were positive for glial fibrillary acidic protein (GFAP). The distribution characteristics of microtubules and intermediate filaments in the two cytoplasm were similar, but the localization, distribution and fluorescence intensity of microfilaments of the two kinds of cells were different. Vimentin-type intermediate filaments were strongly expressed in SHG 44 cells, whereas GFAP was strongly expressed in CHG 5 cells. Acetone, 75% alcohol and 4% paraformaldehyde had a good effect on microtubule fixation. The staining of intermediate filaments (especially Vimentin) with aldehydes was very weak. Conclusion The differentiation of CHG 5 is higher than that of SHG 44, and the difference of cytoskeleton between the two cell lines may be the structural basis of the difference of biological characteristics between the two