Schick B P, Jacoby J A
Cardeza Foundation for Hematologic Research, Department of Medicine, Jefferson Medical College of Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
J Cell Physiol. 1995 Oct;165(1):96-106. doi: 10.1002/jcp.1041650113.
This study has characterized the proteoglycans from the megakaryocytic tumor cell line CHRF 288-11 and the effect of the differentiation-inducing agents phorbol-12-myristate-13-acetate (PMA) and dimethylsulfoxide (DMSO) on proteoglycan synthesis in these cells. There appeared to be two classes of proteoglycans. One, serglycin, was recognized to have a core protein of 31 kDa, an overall molecular mass of 200-300 kDa, and glycosaminoglycan chains of mean size < 25 kDa. The size of this proteoglycan was increased by both PMA and DMSO. Synthesis was increased by PMA and reduced by DMSO. mRNA for serglycin was increased at 24 to 72 hr following PMA treatment. In addition, the cells contained a core protein triplet at 96, 110, and 120 kDa, and the medium only the bands at 96 and 110 kDa, suggesting the presence of betaglycan. Synthesis of this proteoglycan was enhanced by PMA. This proteoglycan had an overall size of 130-150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in control cells, but in the presence of PMA, a component > 250 kDa was present. Probes for Northern blot analysis were prepared by polymerase chain reaction (PCR) based on the sequences of human serglycin and betaglycan. The serglycin probe recognized a 1.4 kb band, and the betaglycan probe recognized a 4.1 kb band, on blots prepared from RNA from CHRF cells and cultured normal human megakaryocytes. Both proteoglycans in their intact form adhered to peptides derived from fibronectin and collagen, but the free GAGs released by alkaline borohydride digestion did not adhere. Synthesis of two proteoglycans appears to be a part of the differentiation process of megakaryocytic tumor cells and normal megakaryocytes.
本研究对巨核细胞肿瘤细胞系CHRF 288-11中的蛋白聚糖进行了表征，并研究了分化诱导剂佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）和二甲基亚砜（DMSO）对这些细胞中蛋白聚糖合成的影响。似乎存在两类蛋白聚糖。一类是丝甘蛋白聚糖，其核心蛋白为31 kDa，总分子量为200 - 300 kDa，糖胺聚糖链平均大小<25 kDa。PMA和DMSO均可增加这种蛋白聚糖的大小。PMA可增加其合成，而DMSO则降低其合成。PMA处理后24至72小时，丝甘蛋白聚糖的mRNA增加。此外，细胞中含有96、110和120 kDa的核心蛋白三联体，而培养基中只有96和110 kDa的条带，提示存在β-聚糖。PMA可增强这种蛋白聚糖的合成。在对照细胞中，这种蛋白聚糖在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上的总大小为130 - 150 kDa，但在PMA存在的情况下，存在一个>250 kDa的组分。基于人丝甘蛋白聚糖和β-聚糖的序列，通过聚合酶链反应（PCR）制备了用于Northern印迹分析的探针。在由CHRF细胞和培养的正常人巨核细胞的RNA制备的印迹上，丝甘蛋白聚糖探针识别出一条1.4 kb的条带，β-聚糖探针识别出一条4.1 kb的条带。两种完整形式的蛋白聚糖均能与源自纤连蛋白和胶原的肽结合，但经碱性硼氢化钠消化释放的游离糖胺聚糖则不能结合。两种蛋白聚糖的合成似乎是巨核细胞肿瘤细胞和正常巨核细胞分化过程的一部分。
