人恶性胶质瘤细胞系CHG-5和SHG-44细胞骨架的比较研究

Comparative study on cytoskeleton of human glioma cell line CHG-5 and SHG-44 in vitro

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 。

Objective To compare the cytoskeleton components between human glioma cell line CHG 5 and SHG 44. Methods The con tents , distribution and fluorescent intensity of microtubules (MTs), microfilaments (MFs) and intermediate filaments (IFs) in the 2 cell lines were detected with immunofluorescence cytochemistry and confocal laser scanning microscopy. The effects of several common fixatives and buffers on every component of cytoskeleton were also studied. Results Besides MTs and MFs, vimentin was expressed in all CHG 5 and SHG 44 cells. Glial fibrillary acid protein (GFAP) positive was only found in a small proportion of the cells. The distributions of MTs and IFs in the cytoplasm were quite similar in both cell lines, but the locations, distributions and fluorescent intensities of MFs were different to some extent. The fluorescent intensity of vimentin was stronger in SHG 44 cells than in CHG 5 cells, and more GFAP positive cells were found in CHG 5 cell line than in SHG 44. The fixatives, 75% ethanol, acetone and 4% paraformaldehyde (4%PFA) gave satisfactory results for MTs immunostaining, but the staining for IFs became weakened or negative after 4% PFA fixation. Conclusion The results suggest that human glioma cell line CHG 5 has a better differentiation than cell line SHG 44. Their differences in biological characteristics might base on their varied states of cytoskeleton.