分子生物学
流式细胞术
细胞生长
细胞因子
白细胞介素2
免疫印迹
受体
生物
白细胞介素10受体,α亚单位
细胞培养
白细胞介素2受体
化学
T细胞
蛋白质亚单位
免疫学
Gα亚单位
免疫系统
基因
生物化学
遗传学
作者
Alicia García de Galdeano,María Dolores Boyano,I Smith-Zubiaga,María Luz Cañavate
出处
期刊:Tumor Biology
[SAGE]
日期:1996-01-01
卷期号:17 (3): 155-167
被引量:21
摘要
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.
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