Piepkorn M, Hovingh P, Linker A
Lab Invest. 1987 Sep;57(3):306-10.
When the glycosaminoglycans (GAGs) of BALB/c 3T3 and B16.F10 melanoma cells were compared during parallel studies, substantial differences were found in the cellular content, rates of turnover, and extent of intrinsic sulfated GAG degradation between the two cell types. The 3T3 cells had more than 10-fold more metabolically-labeled [35S]-GAGs per cell under steady state conditions than did the melanoma cells, and in pulse-chase studies they turned this material over at a much slower rate (apparent respective half-times of 30 hours and 8 hours). Moreover, the labeled material released into the medium during the chase periods was approximately 43% low molecular weight polymeric product, eluting in the included volume of Sephadex G50, for the 3T3 cells, as compared with 79% for the B16.F10 line. For both cell lines approximately 86% of total cell layer-associated polymeric GAG was accessible to GAG-degrading enzymes under conditions which did not lyse the cells, indicating that the majority of cellular GAGs is present external to the plasma membrane. The extensive digestion of sulfated GAGs in evidence for the melanoma cells suggests the involvement of constitutively high cellular levels of GAG-degrading enzymes.
在平行研究中比较BALB/c 3T3细胞和B16.F10黑色素瘤细胞的糖胺聚糖(GAGs)时,发现这两种细胞类型在细胞含量、更新率以及内在硫酸化GAG降解程度方面存在显著差异。在稳态条件下,3T3细胞每个细胞的代谢标记[35S]-GAGs比黑色素瘤细胞多10倍以上,并且在脉冲追踪研究中,它们使这种物质的更新速度要慢得多(表观半衰期分别为30小时和8小时)。此外,在追踪期间释放到培养基中的标记物质,对于3T3细胞来说,约43%是低分子量聚合物产物,在Sephadex G50的包容体积中洗脱,而B16.F10细胞系则为79%。在不裂解细胞的条件下,对于两种细胞系,约86%的与细胞层相关的聚合物GAG可被GAG降解酶作用,这表明大多数细胞GAG存在于质膜外部。黑色素瘤细胞中硫酸化GAG的广泛消化表明细胞中持续高水平的GAG降解酶的参与。