© 1992 by Oxford University Press
Journal of the National Cancer Institute, Vol. 84, No. 3, 165-174,
February 5, 1992
© 1992 Oxford University Press
Coexpression of Vimentin and Keratins by Human Melanoma Tumor Cells: Correlation With Invasive and Metastatic Potential
Department of Anatomy, University of Arizona College of Medicine, and The Arizona Cancer Center Tucson, Ariz
Department of Anatomy, University of Arizona College of Medicine
Department of Pathology, University of Arizona College of Medicine
The Arizona Cancer Center, University of Arizona College of Medicine
Department of Pathology and the University of Arizona College of Medicine, The Arizona Cancer Center
Department of Surgery and the University of Arizona College of Medicine, The Arizona Cancer Center
Department of Hematology/Oncology, University of California Irvine Cancer Center Irvine
Michigan Cancer Foundation Detroit, Mich.
Department of Pathology, Pennsylvania State University School of Medicine Hershey, Pa.
Laboratory of Pathology, Division of Cancer Biology, Diagnosis, and Centers, National Cancer Institute, National Institutes of Health Bethesda, Md.
*Correspondence to: Mary J. C. Hendrix, Ph.D., Department of Anatomy, College of Medicine, University of Arizona, Tucson, AZ 85724.
Background: Several protein markers, including vimentin, have been used to diagnose human melanoma. Because melanoma often has metastasized by the time of diagnosis, early markers prognostic for metastatic potential need to be identified. Commonly, vimentin is found in mesenchymal cells, and keratins are present in epithelial cells, but recent studies report coexpression of vimentin and keratin(s) in epithelial and nonepithelial neoplasms, including some melanomas. Purpose: Our purpose was to determine whether coexpression of vimentin and keratin(s) is correlated with tumor cell invasion and metastatic behavior. Methods: We evaluated nine human melanoma cell lines expressing vimentin and other markers of aggressive tumor behavior (HMB-45, S-100, HLA-ABC class I and HLA-DR class II histocompatibility antigens, and K8 and K18 keratins). Levels of K8 and K18 keratins were determined in the highly metastatic C8161 cell line, the poorly metastatic A375P line, and the moderately metastatic A375M line. To determine whether the presence of keratin affects migratory ability, we altered the conformational structure of keratin filaments in C8161 cells by transfection with a mutant K18 complementary DNA. We also determined messenger RNA levels of human type IV collagenase, an enzyme marker for invasion and metastasis. Results: In A375P cells, two-dimensional electrophoresis with Coomassie-stained gels, immunoblott-ing, and immunofluorescence staining showed no detectable levels of K8 or K18. A375M cells showed low levels of K8 and K18 by Western and Northern blotting, with a distinctive fluorescent subpopulation of cells. In comparison, K8 and K18 levels in C8161 cells were high in all cells. Type IV collagenase messenger RNA levels were lowest in A375P cells and highest in C8161 cells, correlating with invasive ability in vitro and metastatic potential in athymic nude mice. The transfectant clones C1070–10 and C1070–14 derived from the C8161 parent line showed dramatic morphological changes, disrupted keratin filaments, and decreased invasive and metastatic potential directly correlated with a reduction in migratory activity. Conclusion: These findings show a correlation between the coexpression of vimentin with K8 and K18 keratins and the invasive and metastatic behavior of three representative human melanoma cell lines. [J Natl Cancer Inst 84: 165–174, 1992]
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