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大型基质蛋白聚糖、多功能蛋白聚糖和基底膜聚糖由人白血病单核细胞表达并分泌。

Large matrix proteoglycans, versican and perlecan, are expressed and secreted by human leukemic monocytes.

作者信息

Makatsori Evdokia, Lamari Fotini N, Theocharis Achilleas D, Anagnostides Stavros, Hjerpe Anders, Tsegenidis Theodore, Karamanos Nikos K

机构信息

Department of Chemistry, Section of Organic Chemistry, Biochemistry & Natural Products, University of Patras, 26500 Patras, Greece.

出版信息

Anticancer Res. 2003 Jul-Aug;23(4):3303-9.

Abstract

THP-1 is a monocytic cell line originally derived from a patient with acute monocytic leukemia. Interactions of THP-1 cells with other cells and their microenvironment are largely determined by proteoglycans (PGs), the identity of which has not been determined. Previous studies on glycosaminoglycan expression by THP-1 cells and peripheral blood mononuclear cells from healthy individuals showed that both cell types secrete mainly chondroitin sulfate PGs to the culture medium, whereas heparan sulfate PGs are mainly retarded at the cell membrane. However, limited data on the type of PGs synthesized by THP-1 is available. In this study, the identification of PG types synthesised by THP-1 cells, which are not differentiated to macrophages, was examined. Analysis at the mRNA level by RT-PCR showed the expression of six cell membrane-associated PGs: syndecan-1, -2 and -4, glypican-1, thrombomodulin and CD44. Cell extraction, ion-exchange chromatography and dot blot analysis of the isolated PG populations with monoclonal antibodies showed the presence of syndecan-1 and thrombomodulin; the other two syndecans were not detected in any of the isolated populations. The synthesis of matrix PGs was also studied. THP-1 monocytes were positive for the mRNA encoding for versican and perlecan, but not for those encoding for decorin, biglycan, betaglycan and fibromodulin. The mRNA encoding for two versican splice variants V0 (351 bp) and V1 (386 bp), but not for V2, were identified. Biochemical analysis showed the presence of perlecan and of two populations of versican in culture medium with protein cores of average molecular sizes similar to those of V0 and V1. The production of these large matrix PGs by THP-1 monocytes is reported for the first time and may be of importance in monocyte malignant transformation and differentiation.

摘要

THP-1是一种单核细胞系,最初源自一名急性单核细胞白血病患者。THP-1细胞与其他细胞及其微环境的相互作用在很大程度上由蛋白聚糖(PGs)决定,但其具体类型尚未确定。先前对THP-1细胞和健康个体外周血单个核细胞中糖胺聚糖表达的研究表明,这两种细胞类型主要向培养基中分泌硫酸软骨素PGs,而硫酸乙酰肝素PGs主要滞留在细胞膜上。然而,关于THP-1合成的PGs类型的数据有限。在本研究中,对未分化为巨噬细胞的THP-1细胞合成的PGs类型进行了鉴定。通过RT-PCR在mRNA水平进行分析,结果显示六种细胞膜相关PGs的表达:多配体蛋白聚糖-1、-2和-4、磷脂酰肌醇蛋白聚糖-1、血栓调节蛋白和CD44。细胞提取、离子交换色谱以及用单克隆抗体对分离出的PG群体进行斑点印迹分析,结果显示存在多配体蛋白聚糖-1和血栓调节蛋白;在任何分离出的群体中均未检测到其他两种多配体蛋白聚糖。还对基质PGs的合成进行了研究。THP-1单核细胞中编码多功能蛋白聚糖和基底膜聚糖的mRNA呈阳性,但编码核心蛋白聚糖、双糖链蛋白聚糖、β-聚糖和纤调蛋白的mRNA则为阴性。鉴定出了编码多功能蛋白聚糖两种剪接变体V0(351 bp)和V1(386 bp)的mRNA,但未检测到V2的mRNA。生化分析表明,培养基中存在基底膜聚糖和两种多功能蛋白聚糖群体,其蛋白核心的平均分子大小与V0和V1相似。首次报道了THP-1单核细胞产生这些大型基质PGs,这可能在单核细胞恶性转化和分化中具有重要意义。

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