Human melanoma cell lines of primary and metastatic origin express the genes encoding the chains of platelet-derived growth factor (PDGF) and produce a PDGF-like growth factor.
Human melanoma cell lines of primary and metastatic origin express the genes encoding the chains of platelet-derived growth factor (PDGF) and produce a PDGF-like growth factor.
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原发性和转移性人类黑色素瘤细胞系表达编码血小板衍生生长因子 (PDGF) 链的基因，并产生 PDGF 样生长因子。
DOI：
10.1073/pnas.83.19.7197
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发表时间：
1986
影响因子：
11.1
通讯作者：
Koprowski,H
中科院分区：
文献类型：
--
作者：
Westermark,B;Johnsson,A;Paulsson,Y;Betsholtz,C;Heldin,CH;Herlyn,M;Rodeck,U;Koprowski,H
分析正常人黑素细胞和五种人黑色素瘤细胞系的血小板衍生生长因子（PDGF）样活性的产生。三个黑色素瘤细胞系释放的活性，抑制结合125碘标记的PDGF的人包皮成纤维细胞和刺激[3 H]胸苷掺入这些细胞。这些活动被抑制的抗PDGF抗体的加入。所有三种因子产生细胞系均来自同一患者--一种来自原发肿瘤（WM 115），两种来自单个淋巴结转移（WM 239 A和WM 266-4）。对WM 266-4细胞产生的因子进行了详细的生化表征。免疫沉淀，代谢标记的因子迁移在NaDod-SO 4/凝胶电泳作为一个均匀的Mr 31，000的物种，在还原条件下被分解成两个物种的Mr 16，500和Mr 17，000，这意味着分子的二聚体结构。将该因子纯化至均一。通过还原和烷基化因子的反相高压液相色谱分析揭示了与PDGF A链相同的洗脱模式。因此，天然分子似乎是PDGF A链的同源二聚体。用~（32）P标记的PDGF A链和B链（SIS产物）cDNA探针对来自细胞系的poly（A）+ RNA进行印迹杂交分析，揭示了仅来自原发肿瘤组织的细胞系中B链转录物的相对丰度，但A链在所有三种细胞系中表达。我们的结论是，两个结构基因编码的亚基链的PDGF可以在人黑色素瘤细胞中表达，这两个基因可以独立地在这样的细胞中表达。
Normal human melanocytes and five human melanoma cell lines were analyzed for production of platelet-derived growth factor (PDGF)-like activity. Three of the melanoma cell lines released an activity that inhibited binding of 125I-labeled PDGF to human foreskin fibroblasts and stimulated [3H]thymidine incorporation in such cells. These activities were inhibited by the addition of anti-PDGF antibodies. All three factor-producing cell lines were derived from the same patient--one originated from the primary tumor (WM 115), and two were from individual lymph-node metastases (WM 239A and WM 266-4). The factor produced by WM 266-4 cells was characterized biochemically in detail. Immunoprecipitated, the metabolically labeled factor migrated in NaDod-SO4/gel electrophoresis as a homogeneous Mr 31,000 species, which under reducing conditions was resolved into two species of Mr 16,500 and Mr 17,000, implying a dimeric structure of the molecule. The factor was purified to homogeneity. Analysis by reverse-phase high-pressure liquid chromatography of reduced and alkylated factor revealed an elution pattern identical to that of PDGF A chains. Thus, the native molecule appears to be a homodimer of PDGF A chains. Blot-hybridization analysis of poly(A)+ RNA from the cell lines with 32P-labeled PDGF A chain and B chain (SIS product) cDNA probes revealed a relative abundance of B chain transcripts in the cell line originating from the primary tumor tissue only but expression of A chain in all three cell lines. We conclude that the two structural genes encoding each of the subunit chains of PDGF can be expressed in human melanoma cells and that the two genes can be independently expressed in such cells.