title: B16F10 murine melanoma cells express interleukin-2 and a functional interleukin-2 receptor.
abstract: In this study, we analyzed interleukin-2 (IL-2) and IL-2 receptor (IL-2R) expression in murine B16F10 melanoma and studied the effect of recombinant IL-2 (rIL-2) on the proliferation of these cells. Flow cytometry analysis revealed the presence of the IL-2R alpha subunit in B16F10 melanoma, with a mean positivity rate of 30%. Using confocal microscopy, the expression of this chain could be visualized on the surface of B16F10 cells and in intracellular compartments when the cells were permeabilized with ethanol. In addition to the alpha subunit, the IL-2R beta subunit was also expressed in B16F10 cells as shown by reverse transcription and polymerase chain reaction analysis. The functionality of the IL-2R on B16F10 cells was shown by the fact that cell proliferation increased dose-dependently with the addition of rIL-2 to the culture medium. We also detected expression of the IL-2 gene in B16F10 cells. In Northern blot assays, a typical band of 0.9 kb corresponding to IL-2 mRNA was observed, although supernatants from B16F10 cultures had no detectable IL-2 activity. Furthermore, the addition of neutralizing antibody (anti-IL-2) to cell cultures had no effect on cell proliferation. From these results, we concluded that an IL-2 signalling system is present in murine B16F10 melanoma cells and that IL-2 favors B16F10 cell proliferation, suggesting a role for this cytokine in the tumoral activity of these cells.
authors.affiliation: Department of Cell Biology and Morphological Sciences, School of Medicine, University of the Basque Country, Leioa, Spain.
authors.forename: A
authors.initials: A
authors.lastname: García de Galdeano
authors.forename: M D
authors.initials: MD
authors.lastname: Boyano
authors.forename: I
authors.initials: I
authors.lastname: Smith-Zubiaga
authors.forename: M L
authors.initials: ML
authors.lastname: Cañavate
mesh_terms: D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D017931:DNA Primers / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D007376:Interleukin-2 / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D012334:RNA, Neoplasm / Q000235:genetics; D015375:Receptors, Interleukin-2 / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D007376:Interleukin-2; D012334:RNA, Neoplasm; D015375:Receptors, Interleukin-2