Characterization of extracellular matrix-associated glycosaminoglycans produced by untransformed and transformed bovine corneal endothelial cells in culture.
Characterization of extracellular matrix-associated glycosaminoglycans produced by untransformed and transformed bovine corneal endothelial cells in culture.
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培养中未转化和转化的牛角膜内皮细胞产生的细胞外基质相关糖胺聚糖的表征。
DOI：
10.1111/j.1432-1033.1985.tb09277.x
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Nicolson,GL
中科院分区：
文献类型：
--
作者：
Wang,ZW;Irimura,T;Nakajima,M;Belloni,PN;Nicolson,GL
用猿猴病毒 40 体外转化克隆的牛角膜内皮细胞系，分离细胞合成的内皮下细胞外基质相关的硫酸化糖胺聚糖，并与未转化的对应物进行比较。在不存在成纤维细胞生长因子的情况下，转化的内皮细胞比未转化的细胞以更快的速率生长，达到更高的静止细胞密度。在每个细胞的基础上，转化细胞产生的硫酸化糖胺聚糖的量略低。在培养 7 天期间，细胞外基质中硫酸化糖胺聚糖的产生率增加。在汇合时，由未转化的内皮细胞合成的与细胞外基质相关的硫酸化糖胺聚糖由约80%的硫酸乙酰肝素和约20%的硫酸软骨素组成。转化内皮细胞的细胞外基质相关硫酸化糖胺聚糖由约 70% 硫酸乙酰肝素和约 30% 硫酸软骨素加硫酸皮肤素组成。来自未转化和转化内皮细胞的基质相关硫酸乙酰肝素在 Fractogel TSK HW-55(S) 上的高速凝胶渗透色谱图非常相似，并给出单峰 (Kav= 0.19)。根据峰的洗脱位置估计，表观 M 约为 47000。来自未转化和转化内皮细胞的硫酸乙酰肝素通过与含有乙酰肝素酶（硫酸乙酰肝素特异性内切 β-葡萄糖醛酸酶）的转移性 B16 黑色素瘤细胞裂解液一起孵育而被降解。硫酸乙酰肝素降解产物在凝胶渗透柱上的洗脱位置相似(Kav=0.43)。低pH下亚硝酸脱氨基后的降解产物的尺寸分析和阴离子交换色谱表明，硫酸乙酰肝素在未转化和转化的内皮细胞中的N-硫酸化程度相似。结果表明，内皮细胞的转化仅轻微改变内皮下基质相关硫酸化糖胺聚糖的分子性质。
A cloned bovine corneal endothelial cell line was transformedin vitroby simian virus 40, and the subendothelial extracellular matrix‐associated sulfated glycosaminoglycans synthesized by the cells were isolated and compared with their untransformed counterpart. The transformed endothelial cells grew at faster rates to higher stationary cell densities in the absence of fibroblast growth factor than did the untransformed cells. On a per‐cell basis, the transformed cells produced slightly lower amounts of sulfated glycosaminoglycans. The rate of production of sulfated glycosaminoglycans in extracellular matrix increased during seven days of culture. At confluency the extracellular matrix‐associated sulfated glycosaminoglycans synthesized by the untransformed endothelial cells consisted of about 80% heparan sulfate and about 20% chondroitin sulfate. Extracellular matrix‐associated sulfated glycosaminoglycans of transformed endothelial cells were composed of about 70% heparan sulfate and about 30% chondroitin sulfate plus dermatan sulfate. High‐speed gel permeation chromatography profiles on Fractogel TSK HW‐55(S) of matrix‐associated heparan sulfate from untransformed and transformed endothelial cells were very similar, and gave single peaks (Kav= 0.19). Apparent Mrestimated from the eluting position of the peaks were approximately 47000. Heparan sulfate from both untransformed and transformed endothelial cells was degraded by incubation with a metastatic B16 melanoma cell lysate containing heparanase (heparan‐sulfate‐specific endo‐β‐glucuronidase). The eluting position of the heparan sulfate degradation products on gel permeation column were similar (Kav= 0.43). Size analysis and anion‐exchange chromatography of the degradation products after nitrous acid deamination at low pH indicated that the degree ofN‐sulfation of heparan sulfate was similar in untransformed and transformed endothelial cells. The results indicated that transformation of endothelial cells only slightly changes the molecular nature of subendothelial matrix‐associated sulfated glycosaminoglycans.