Abstract
The effects of IFN-gamma and interleukin 4 (IL-4) on cell proliferation and two-dimensional gel electrophoretic protein patterns of the human renal carcinoma cell line ACHN were studied. Treatment of the cells with IFN-gamma resulted in a 40-50% decrease in their proliferation. IL-4 treatment resulted in a 30-40% decrease. Treating cells with both cytokines had the same effect as with IFN-gamma alone, thus precluding a synergistic antiproliferative interaction of these two cytokines. To identify IL-4- and IFN-gamma-regulated proteins in ACHN, two-dimensional preparative gel electrophoresis was used, combined with either capillary electrophoresis or high-performance liquid chromatography and either Edman or mass spectrometric sequencing. The following cytokine-induced proteins were identified: tropomyosin, heat shock protein 27, manganese superoxide dismutase, glutathione S-transferase pi, and protein kinase C inhibitor I. Tropomyosin increased 2-fold when cells were treated with IFN-gamma. Levels of heat shock protein 27 increased 2-fold with IL-4, 3-fold with IFN-gamma, and 4-fold when the cytokines were used in combination. Manganese superoxide dismutase increased 3-fold with IFN-gamma but was unaffected by IL-4. Glutathione S-transferase pi increased 3-fold with IFN-gamma. Levels of protein kinase C inhibitor I increased greater than 3-fold with IL-4, 4-fold with IFN-gamma, and 7-fold when both cytokines were used. In addition, the following constitutive ACHN proteins were identified: copper zinc superoxide dismutase, 60S acidic ribosomal protein P2, and a second heat shock protein 27 isoform. These findings help define the biochemical modes of action of IFN-gamma and IL-4 and their potential in the biological therapy of renal cell carcinoma.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Carcinoma, Renal Cell / chemistry
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Carcinoma, Renal Cell / pathology*
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Cell Division / drug effects
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Chromatography, High Pressure Liquid
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Drug Synergism
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Electrophoresis, Gel, Two-Dimensional*
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Gene Expression Regulation, Neoplastic / drug effects*
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Glutathione S-Transferase pi
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Glutathione Transferase / analysis
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Glutathione Transferase / biosynthesis
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Glutathione Transferase / genetics
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Growth Inhibitors / pharmacology*
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Heat-Shock Proteins / analysis
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Heat-Shock Proteins / biosynthesis
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Heat-Shock Proteins / genetics
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Humans
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Interferon-gamma / pharmacology*
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Interleukin-4 / pharmacology*
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Isoenzymes / analysis
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Isoenzymes / biosynthesis
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Isoenzymes / genetics
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Kidney Neoplasms / chemistry
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Kidney Neoplasms / pathology*
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Mass Spectrometry
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Molecular Sequence Data
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Neoplasm Proteins / analysis
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Neoplasm Proteins / biosynthesis*
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Neoplasm Proteins / genetics
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Nerve Tissue Proteins / analysis
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Nerve Tissue Proteins / biosynthesis
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Nerve Tissue Proteins / genetics
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Phosphoproteins / analysis
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Phosphoproteins / biosynthesis
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Phosphoproteins / genetics
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Recombinant Proteins / pharmacology
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Ribosomal Proteins
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Sequence Analysis
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Superoxide Dismutase / analysis
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Superoxide Dismutase / biosynthesis
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Superoxide Dismutase / genetics
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Tropomyosin / analysis
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Tropomyosin / biosynthesis
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Tropomyosin / genetics
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Tumor Cells, Cultured / drug effects
Substances
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Growth Inhibitors
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HINT1 protein, human
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Heat-Shock Proteins
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Isoenzymes
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Neoplasm Proteins
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Nerve Tissue Proteins
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Phosphoproteins
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Recombinant Proteins
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Ribosomal Proteins
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Tropomyosin
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phosphoprotein P2, ribosomal
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Interleukin-4
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Interferon-gamma
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Superoxide Dismutase
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GSTP1 protein, human
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Glutathione S-Transferase pi
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Glutathione Transferase