Chandrasekaran E V, Davidson E A
Cancer Res. 1979 Mar;39(3):870-80.
Glycosaminoglycans have been characterized from a normal human breast cell line (HBL-100) and two different cell lines from human breast carcinoma (MDA-MB-231 and MCF-7). The glycosaminoglycans were labeled by exposure of cell cultures to [3H]glucosamine and [35S]sulfate and then isolated from both spent media and cells by pronase digestion and cetylpyridinium chloride fractionation. They were further characterized by (a) hexosamine composition, (b) controlled-pore glass exclusion chromatography, (c) reactivity with specific enzymes (hyaluronidase chondroitinase, heparitinase, and heparinase), (d) nitrous acid degradation, and (e) DEAD-Sephadex chromatography. The results indicate that the HBL-100 line synthesizes mainly hyaluronic acid, most of which is secreted into the medium. Chondroitin sulfate and heparan sulfate are the predominant glycosaminoglycans synthesized by the cancer lines; both are found mainly in the spent medium, but the hyaluronic acid synthesized by the MDA-MB-231 line remains cell associated. The cell-associated heparan sulfate had a molecular weight in excess of 13,000 and may contain linkages susceptible to testicular hyaluronidase. The MCF-7 cells produce significantly lower amounts of glycosaminoglycans than do the other two lines.
已对来自正常人乳腺细胞系(HBL - 100)以及两种不同的人乳腺癌细胞系(MDA - MB - 231和MCF - 7)的糖胺聚糖进行了表征。通过将细胞培养物暴露于[3H]葡糖胺和[35S]硫酸盐来标记糖胺聚糖,然后通过链霉蛋白酶消化和十六烷基吡啶氯化物分级分离从用过的培养基和细胞中分离出来。通过以下方法对它们进行进一步表征:(a)己糖胺组成,(b)可控孔径玻璃排阻色谱法,(c)与特定酶(透明质酸酶、软骨素酶、类肝素酶和肝素酶)的反应性,(d)亚硝酸降解,以及(e)DEAE - 葡聚糖凝胶色谱法。结果表明,HBL - 100细胞系主要合成透明质酸,其中大部分分泌到培养基中。硫酸软骨素和硫酸乙酰肝素是癌细胞系合成的主要糖胺聚糖;两者主要存在于用过的培养基中,但MDA - MB - 231细胞系合成的透明质酸仍与细胞相关。与细胞相关的硫酸乙酰肝素分子量超过13,000,可能含有易被睾丸透明质酸酶作用的连接键。MCF - 7细胞产生的糖胺聚糖量明显低于其他两个细胞系。