Home LiteratureArticle Details
PMID: 3020539 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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.

Westermark B, Johnsson A, Paulsson Y, Betsholtz C, Heldin CH, Herlyn M, Rodeck U, Koprowski H

Abstract

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.

MeSH Terms
Cell Line DNA, Neoplasm/genetics Gene Expression Regulation Humans Macromolecular Substances Melanoma, Experimental/genetics Neoplasm Metastasis Platelet-Derived Growth Factor/genetics Proto-Oncogene Proteins/genetics RNA, Messenger/genetics Receptors, Cell Surface/metabolism Receptors, Platelet-Derived Growth Factor
Chemicals
DNA, Neoplasm Macromolecular Substances Platelet-Derived Growth Factor Proto-Oncogene Proteins RNA, Messenger Receptors, Cell Surface Receptors, Platelet-Derived Growth Factor
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Westermark B
Johnsson A
Paulsson Y
Betsholtz C
Heldin C H
Herlyn M
Rodeck U
Koprowski H
References (33)
33 references, click to expand
  1. Cellular genes analogous to retroviral onc genes are transcribed in human tumour cells.
    Nature. 1982 Jan 14;295(5845):116-9 PMID: 6173755
  2. Demonstration of an antibody against platelet-derived growth factor.
    Exp Cell Res. 1981 Dec;136(2):255-61 PMID: 6171440
  3. Nucleotide sequence of the simian sarcoma virus genome: demonstration that its acquired cellular sequences encode the transforming gene product p28sis.
    Proc Natl Acad Sci U S A. 1983 Feb;80(3):731-5 PMID: 6298772
  4. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor.
    Science. 1983 Jul 15;221(4607):275-7 PMID: 6304883
  5. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus.
    Nature. 1983 Jul 7-13;304(5921):35-9 PMID: 6306471
  6. Expression of a platelet-derived growth factor-like protein in simian sarcoma virus transformed cells.
    Science. 1983 Sep 30;221(4618):1348-50 PMID: 6310754
  7. Structural and immunological similarities between simian sarcoma virus gene product(s) and human platelet-derived growth factor.
    Nature. 1983 Oct 13-19;305(5935):605-8 PMID: 6312326
  8. Human-proto-oncogene nucleotide sequences corresponding to the transforming region of simian sarcoma virus.
    Science. 1984 Feb 3;223(4635):487-91 PMID: 6318322
  9. Synthesis of a PDGF-like growth factor in human glioma and sarcoma cells suggests the expression of the cellular homologue to the transforming protein of simian sarcoma virus.
    Biochem Biophys Res Commun. 1983 Nov 30;117(1):176-82 PMID: 6318746
  10. Growth factor-induced proliferation of human fibroblasts in serum-free culture depends on cell density and extracellular calcium concentration.
    J Cell Physiol. 1984 Feb;118(2):203-10 PMID: 6607257
  11. A glioma-derived analog to platelet-derived growth factor: demonstration of receptor competing activity and immunological crossreactivity.
    Proc Natl Acad Sci U S A. 1984 Feb;81(3):926-30 PMID: 6322178
  12. Transformation of NIH 3T3 cells by a human c-sis cDNA clone.
    Nature. 1984 Mar 29-Apr 4;308(5958):464-7 PMID: 6323994
  13. Nucleotide sequence analysis identifies the human c-sis proto-oncogene as a structural gene for platelet-derived growth factor.
    Cell. 1984 May;37(1):123-9 PMID: 6327048
  14. The c-sis gene encodes a precursor of the B chain of platelet-derived growth factor.
    EMBO J. 1984 May;3(5):921-8 PMID: 6329745
  15. Expression of the normal human sis/PDGF-2 coding sequence induces cellular transformation.
    Cell. 1984 Nov;39(1):89-97 PMID: 6091919
  16. A study of tumor progression: the precursor lesions of superficial spreading and nodular melanoma.
    Hum Pathol. 1984 Dec;15(12):1147-65 PMID: 6500548
  17. Purification and properties of porcine platelet-derived growth factor.
    EMBO J. 1984 Dec 1;3(12):2963-7 PMID: 6526008
  18. Primary melanoma cells of the vertical growth phase: similarities to metastatic cells.
    J Natl Cancer Inst. 1985 Feb;74(2):283-9 PMID: 3856042
  19. Platelet-derived growth factor.
    Mol Cell Endocrinol. 1985 Mar;39(3):169-87 PMID: 2984061
  20. Synthesis and secretion of proteins resembling platelet-derived growth factor by human glioblastoma and fibrosarcoma cells in culture.
    Proc Natl Acad Sci U S A. 1985 Apr;82(8):2404-8 PMID: 3857590
  21. Production of PDGF-like growth factor by breast cancer cell lines.
    Int J Cancer. 1985 Aug 15;36(2):247-52 PMID: 2991148
  22. Cultured human endothelial cells express platelet-derived growth factor B chain: cDNA cloning and structural analysis.
    Nature. 1985 Aug 22-28;316(6030):748-50 PMID: 4033772
  23. Antibodies against platelet-derived growth factor inhibit acute transformation by simian sarcoma virus.
    Nature. 1985 Oct 3-9;317(6036):438-40 PMID: 2995828
  24. Characteristics of cultured human melanocytes isolated from different stages of tumor progression.
    Cancer Res. 1985 Nov;45(11 Pt 2):5670-6 PMID: 4053039
  25. A human osteosarcoma cell line secretes a growth factor structurally related to a homodimer of PDGF A-chains.
    Nature. 1986 Feb 6-12;319(6053):511-4 PMID: 3456080
  26. cDNA sequence and chromosomal localization of human platelet-derived growth factor A-chain and its expression in tumour cell lines.
    Nature. 1986 Apr 24-30;320(6064):695-9 PMID: 3754619
  27. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent.
    Biochem J. 1973 Jul;133(3):529-39 PMID: 4733239
  28. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma.
    J Cell Biol. 1975 Dec;67(3):835-51 PMID: 811671
  29. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film.
    FEBS Lett. 1977 Oct 15;82(2):314-6 PMID: 913604
  30. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA.
    Eur J Biochem. 1980 Jun;107(2):303-14 PMID: 6772444
  31. Chemical and biological properties of a growth factor from human-cultured osteosarcoma cells: resemblance with platelet-derived growth factor.
    J Cell Physiol. 1980 Nov;105(2):235-46 PMID: 6936400
  32. Specific receptors for platelet-derived growth factor on cells derived from connective tissue and glia.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3664-8 PMID: 6943569
  33. Platelet-derived growth factor: identification of constituent polypeptide chains.
    Biochem Biophys Res Commun. 1982 Jan 15;104(1):66-74 PMID: 7073684
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1986-10-00
Pages
7197-200
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386682
Subset
IM
Grants
NCI NIH HHS · CA-10815 · United States
NCI NIH HHS · CA-21124 · United States
NCI NIH HHS · CA-25874 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com