Edward M
J Cell Sci. 1986 Sep;85:217-29. doi: 10.1242/jcs.85.1.217.
The effect of ascorbate on the glycosaminoglycans synthesized by normal and simian virus 40(SV40)-transformed human skin fibroblasts was examined. Cells were incubated in the presence or absence of ascorbate, and radiolabelled with [3H]glucosamine and [35S]sulphate for 48 h, 3 days after reaching confluence. Glycosaminoglycans were analysed in the medium, a collagenase extract, and in the trypsin/cell-associated fraction. Hyaluronic acid was the main 3H-labelled glycosaminoglycan in all but the collagenase extracts, and showed a large decrease in normal fibroblast cultures, but a significant increase in SV40-transformed fibroblast cultures following feeding with ascorbate. Incorporation of [3H]glucosamine into sulphated glycosaminoglycans was reduced in normal fibroblast cultures but increased slightly in SV40-transformed cultures following ascorbate supplementation. [35S]sulphate incorporation remained essentially unaltered in both cell cultures. Ascorbate stimulated the deposition of glycosaminoglycans into the insoluble matrix of normal fibroblasts while reducing the deposition in SV40-transformed fibroblast cultures. The observed changes may in part be related to ascorbate-induced deposition of collagen in normal fibroblast cultures and the inability of the transformed fibroblast cells to deposit an extensive extracellular matrix, in addition to possible changes in the specific activity of the UDP-N-acetyl-[3H]hexosamine pool.
研究了抗坏血酸对正常及猿猴病毒40（SV40）转化的人皮肤成纤维细胞合成糖胺聚糖的影响。细胞在有或没有抗坏血酸的情况下孵育，并在汇合后3天用[3H]葡糖胺和[35S]硫酸盐进行放射性标记48小时。对培养基、胶原酶提取物以及胰蛋白酶/细胞相关部分中的糖胺聚糖进行分析。除胶原酶提取物外，透明质酸是所有样本中主要的3H标记糖胺聚糖，在正常成纤维细胞培养物中显著减少，但在抗坏血酸处理后的SV40转化成纤维细胞培养物中显著增加。在正常成纤维细胞培养物中，[3H]葡糖胺掺入硫酸化糖胺聚糖的量减少，但在抗坏血酸补充后的SV40转化培养物中略有增加。在两种细胞培养物中，[35S]硫酸盐掺入量基本保持不变。抗坏血酸刺激糖胺聚糖在正常成纤维细胞的不溶性基质中沉积，同时减少在SV40转化成纤维细胞培养物中的沉积。观察到的变化可能部分与抗坏血酸诱导的正常成纤维细胞培养物中胶原蛋白的沉积以及转化的成纤维细胞无法沉积广泛的细胞外基质有关，此外还可能与UDP-N-乙酰-[3H]己糖胺池的比活性变化有关。
