Piepkorn M, Carney H, Linker A
Lab Invest. 1985 Aug;53(2):187-93.
Glycosaminoglycan synthesis was studied in cell populations of ultraviolet light-induced murine cutaneous fibrosarcoma cells under conditions of varying growth rates in vitro. After labeling with the precursors, 3H-glucosamine and 35SO4, sulfated glycosaminoglycans recoverable by direct proteolysis of the culture monolayers increased approximately 5-fold on a per cell basis from sparsely populated, exponential cell cultures (greater than 85% of cells in S, G2, or M phases) to stationary cultures inhibited by high cell density (greater than 50% of cells in G1). Within this cell surface-associated material, the relative ratio of heparan sulfate to the chondroitin sulfates was approximately 60/40% under conditions of exponential growth; in the growth-arrested cultures, the reverse ratio was found. The substratum attached material, obtained from the flask surface after ethyl glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA)-mediated detachment of the monolayers, contained relatively more hyaluronic acid, heparan sulfate, and chondroitin sulfates in the most actively proliferating cultures compared with the growth-inhibited cell populations. Furthermore, heparan sulfate and the chondroitin sulfates, which were enriched in the substratum material and in the cell pellet of exponential cultures, showed a relative shift to the cell surface-associated compartment (releasable by mild trypsinization after EGTA-mediated cell detachment) and to the compartment loosely associated with the pericellular matrix (i.e., released into the supernatant during detachment of the monolayers in the presence of EGTA). These results demonstrate that a variety of differences in the quantities, relative compositional ratios, and cell compartment distributions of hyaluronic acid and sulfated glycosaminoglycans occur in fibrosarcoma cell populations which vary in their rate of cell growth consequent to cell density in culture.
在体外不同生长速率条件下，对紫外线诱导的小鼠皮肤纤维肉瘤细胞群体中的糖胺聚糖合成进行了研究。在用前体3H-葡萄糖胺和35SO4标记后，通过对培养单层进行直接蛋白水解可回收的硫酸化糖胺聚糖，以每细胞为基础，从稀疏接种的指数生长细胞培养物（处于S期、G2期或M期的细胞大于85%）到受高细胞密度抑制的静止培养物（处于G1期的细胞大于50%）增加了约5倍。在这种与细胞表面相关的物质中，硫酸乙酰肝素与硫酸软骨素的相对比例在指数生长条件下约为60/40%；在生长停滞的培养物中，发现比例相反。从乙二醇双（β-氨基乙醚）-N，N'-四乙酸（EGTA）介导的单层细胞脱离后从培养瓶表面获得的附着于基质的物质，与生长受抑制的细胞群体相比，在增殖最活跃的培养物中含有相对更多的透明质酸、硫酸乙酰肝素和硫酸软骨素。此外，在指数培养物的基质物质和细胞沉淀中富集的硫酸乙酰肝素和硫酸软骨素，显示出相对转移到与细胞表面相关的区室（在EGTA介导的细胞脱离后经轻度胰蛋白酶消化可释放）和与细胞周基质松散相关的区室（即在EGTA存在下单层细胞脱离期间释放到上清液中）。这些结果表明，在因培养中的细胞密度而细胞生长速率不同的纤维肉瘤细胞群体中，透明质酸和硫酸化糖胺聚糖在数量、相对组成比例和细胞区室分布方面存在多种差异。
