Sullivan C M, Smith D M, Matsui N M, Andrews L E, Clauser K R, Chapeaurouge A, Burlingame A L, Epstein L B
Cancer Research Institute, Department of Pediatrics, University of California-San Francisco, 94143, USA.
Cancer Res. 1997 Mar 15;57(6):1137-43.
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.
研究了γ-干扰素（IFN-γ）和白细胞介素4（IL-4）对人肾癌细胞系ACHN细胞增殖及二维凝胶电泳蛋白质图谱的影响。用IFN-γ处理细胞导致其增殖下降40%-50%。用IL-4处理导致增殖下降30%-40%。用两种细胞因子处理细胞与单独用IFN-γ处理效果相同，因此排除了这两种细胞因子的协同抗增殖相互作用。为了鉴定ACHN细胞中IL-4和IFN-γ调节的蛋白质，采用了二维制备凝胶电泳，并结合毛细管电泳或高效液相色谱以及埃德曼测序或质谱测序。鉴定出了以下细胞因子诱导的蛋白质：原肌球蛋白、热休克蛋白27、锰超氧化物歧化酶、谷胱甘肽S-转移酶π和蛋白激酶C抑制剂I。当细胞用IFN-γ处理时，原肌球蛋白增加了2倍。热休克蛋白27的水平在IL-4处理时增加2倍，在IFN-γ处理时增加3倍，在两种细胞因子联合使用时增加4倍。锰超氧化物歧化酶在IFN-γ处理时增加3倍，但不受IL-4影响。谷胱甘肽S-转移酶π在IFN-γ处理时增加3倍。蛋白激酶C抑制剂I的水平在IL-4处理时增加超过3倍，在IFN-γ处理时增加4倍，在两种细胞因子都使用时增加7倍。此外，还鉴定出了以下ACHN组成性蛋白质：铜锌超氧化物歧化酶、60S酸性核糖体蛋白P2和另一种热休克蛋白27异构体。这些发现有助于确定IFN-γ和IL-4的生化作用模式及其在肾细胞癌生物治疗中的潜力。
