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-Interferon- and Interleukin 4-regulated Proteins in the Human Renal Carcinoma Cell Line ACHN1
Cancer Research Institute [C. M. S., D. M. S., N. M. M., L. B. E.], Departments of Pediatrics [C. M. S., N. M. M., L. B. E.] and Pharmaceutical Chemistry [L. E. A., K. R. C., A. C., A. L. B.], and Liver Center [A. L. B.], University of California-San Francisco, San Francisco, California 94143
The effects of IFN-
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-
resulted in a 4050% decrease in their proliferation. IL-4 treatment resulted in a 3040% decrease. Treating cells with both cytokines had the same effect as with IFN-
alone, thus precluding a synergistic antiproliferative interaction of these two cytokines. To identify IL-4- and IFN-
-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
, and protein kinase C inhibitor I. Tropomyosin increased 2-fold when cells were treated with IFN-
. Levels of heat shock protein 27 increased 2-fold with IL-4, 3-fold with IFN-
, and 4-fold when the cytokines were used in combination. Manganese superoxide dismutase increased 3-fold with IFN-
but was unaffected by IL-4. Glutathione S-transferase
increased 3-fold with IFN-
. Levels of protein kinase C inhibitor I increased greater than 3-fold with IL-4, 4-fold with IFN-
, 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-
and IL-4 and their potential in the biological therapy of renal cell carcinoma.
1 This work was supported by NIH Grants CA 27903 and CA 44446 and a University of California Systemwide Biotechnology Grant (to L. B. E.) and by NIH Grants DK266743, RR01614, and ES04705 and NSF Grant DIR8700766 (to A. L. B.). A portion of this work was presented at the 87th Annual Meeting of the American Association for Cancer Research, April 20, 1996, Washington, DC.
2 Present address: Center for Biomedical Laboratory Science, San Francisco State University, San Francisco, CA 94132.
3 To whom requests for reprints should be addressed, at Box 0748, Department of Pediatrics, University of California-San Francisco, San Francisco, CA 94143; Phone: 415-476-2981; Fax: 415-476-9976.
Received 10/ 4/96. Accepted 1/18/97.
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