Toda M, Miura M, Asou H, Toya S, Uyemura K
Department of Physiology, Keio University, School of Medicine, Tokyo, Japan.
J Neurochem. 1994 Nov;63(5):1975-8. doi: 10.1046/j.1471-4159.1994.63051975.x.
The cellular functions of the intermediate filament family including glial fibrillary acidic protein (GFAP) are not well known yet beyond their roles as structural elements of cells. Expression of GFAP, which is specific in astrocytes and regulated developmentally, suggests its involvement in cell growth and differentiation of astrocytes. We transfected murine GFAP cDNA into a rat astrocytoma C6 cell line to assess the specific effect of GFAP on cells. Two stable GFAP-transfected cell lines, GFC6-5 and GFC6-6, exhibited a series of morphological and growth characteristics that distinguish them from their counterparts, i.e., NeoC6 cells transfected only with the neomycin-resistant gene, and native C6 cells. Both GFC6-5 and GFC6-6 cells showed elongated cell shapes with extended processes rich in GFAP, markedly suppressed cell growth, and decreased bromodeoxyuridine uptake. Western blot analysis revealed a remarkable increase of GFAP expression in GFC6-5 and GFC6-6 compared with that in NeoC6 and C6, in contrast to similar vimentin expression in all cell lines. The results indicate that the expression of GFAP has dramatic effects on cell morphology and cell growth suppression in C6 cells, suggesting that GFAP may function as a tumor suppressor in astrocytoma.
包括胶质纤维酸性蛋白(GFAP)在内的中间丝家族的细胞功能,除了作为细胞的结构成分外,目前还不太清楚。GFAP在星形胶质细胞中具有特异性表达且受发育调控,这表明它参与了星形胶质细胞的生长和分化。我们将小鼠GFAP cDNA转染到大鼠星形细胞瘤C6细胞系中,以评估GFAP对细胞的特定作用。两个稳定转染GFAP的细胞系GFC6 - 5和GFC6 - 6,表现出一系列形态和生长特征,使其与对照细胞有所区别,即仅转染了新霉素抗性基因的NeoC6细胞和天然C6细胞。GFC6 - 5和GFC6 - 6细胞均呈现出细长的细胞形态,具有富含GFAP的延长突起,细胞生长明显受到抑制,溴脱氧尿苷摄取减少。蛋白质免疫印迹分析显示,与NeoC6和C6细胞相比,GFC6 - 5和GFC6 - 6细胞中GFAP表达显著增加,而所有细胞系中的波形蛋白表达相似。结果表明,GFAP的表达对C6细胞的形态和细胞生长抑制具有显著影响,提示GFAP可能在星形细胞瘤中发挥肿瘤抑制作用。