Molecular cloning of a human melanoma-associated chondroitin sulfate proteoglycan
Molecular cloning of a human melanoma-associated chondroitin sulfate proteoglycan
复制标题
DOI：
10.1073/pnas.93.18.9710
复制
发表时间：
1996-09-03
影响因子：
11.1
通讯作者：
Reisfeld, RA
中科院分区：
文献类型：
--
作者：
Pluschke, G;Vanek, M;Reisfeld, RA
人黑色素瘤相关硫酸软骨素蛋白多糖(MCSP)被mAb9.2.27识别，在黑色素瘤细胞在内皮基底膜上扩散的早期事件中起到稳定细胞-基质相互作用的作用。我们报道了编码人MCSP整个核心蛋白的cDNA的分子克隆和核苷酸测序，并给出了其推导的氨基酸序列，该核心蛋白包含一个2322aa的开放阅读框，包括一个较大的胞外结构域，一个疏水性的跨膜区，以及一个相对较短的细胞质尾部。Northern杂交分析表明，MCSP cDNA探针检测到单个8.0kb的RNA在人黑色素瘤细胞系中表达，并与MCSP编码序列的一段片段进行了原位杂交实验，定位于黑色素瘤皮肤转移组织中的MCSP mRNA。用MCSP特异的探针与人的多个Northern杂交显示，MCSP只与黑色素瘤细胞杂交，而不与其他人类癌细胞或各种人胎儿和成人组织杂交。这些数据表明，MCSP代表由人恶性黑色素瘤细胞表达的一种完整的膜硫酸软骨素蛋白多糖，编码MCSP的cDNAs的存在将有助于研究该分子的结构和功能之间的关系，并有助于确定其在人类恶性黑色素瘤发生发展中的作用。
A human melanoma-associated chondroitin sulfate proteoglycan (MCSP), recognized by mAb 9.2.27, plays a role in stabilizing cell-substratum interactions during early events of melanoma cell spreading on endothelial basement membranes. We report here the molecular cloning and nucleotide sequencing of cDNA encoding the entire core protein of human MCSP and provide its deduced amino acid sequence, This core protein contains an open reading frame of 2322 aa, encompassing a large extracellular domain, a hydrophobic transmembrane region, and a relatively short cytoplasmic tail, Northern blot analysis indicated that MCSP cDNA probes detect a single 8.0-kb RNA species expressed in human melanoma cell lines, In situ hybridization experiments with a segment of the MCSP coding sequence localized MCSP mRNA in biopsies prepared from melanoma skin metastases. Multiple human Northern blots with an MCSP-specific probe revealed a strong hybridization signal only with melanoma cells and not with other human cancer cells or a variety of human fetal and adult tissues, These data indicate that MCSP represents an integral membrane chondroitin sulfate proteoglycan expressed by human malignant melanoma cells, The availability of cDNAs encoding MCSP should facilitate studies designed to establish correlations between structure and function of this molecule and help to establish its role in the progression of human malignant melanoma.