Identification of constitutive and gamma-interferon- and interleukin 4-regulated proteins in the human renal carcinoma cell line ACHN.
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.
Sullivan
C M
CM
Cancer Research Institute, Department of Pediatrics, University of California-San Francisco, 94143, USA.
Smith
D M
DM
Matsui
N M
NM
Andrews
L E
LE
Clauser
K R
KR
Chapeaurouge
A
A
Burlingame
A L
AL
Epstein
L B
LB
D000595:Amino Acid Sequence; D002292:Carcinoma, Renal Cell / Q000737:chemistry / Q000473:pathology*; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D004357:Drug Synergism; D015180:Electrophoresis, Gel, Two-Dimensional; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D006131:Growth Inhibitors / Q000494:pharmacology*; D006360:Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D015847:Interleukin-4 / Q000494:pharmacology*; D007527:Isoenzymes / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D007680:Kidney Neoplasms / Q000737:chemistry / Q000473:pathology*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D010750:Phosphoproteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D012269:Ribosomal Proteins; D017421:Sequence Analysis; D013482:Superoxide Dismutase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D014335:Tropomyosin / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured / Q000187:drug effects
D006131:Growth Inhibitors; C063306:HINT1 protein, human; D006360:Heat-Shock Proteins; D007527:Isoenzymes; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; D014335:Tropomyosin; C031937:phosphoprotein P2, ribosomal; D015847:Interleukin-4; D007371:Interferon-gamma; D013482:Superoxide Dismutase; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase