Abstract
Interleukin (IL-2) and IL-2Rβ/γ have been shown to be expressed in human carcinomas in culture and in situ. Recently, expression of endogenous IL-2 and IL-2R in the cytoplasm was found to be up-regulated in tumour cells undergoing mitosis. This observation suggested that similar to its role in lymphocytes, the IL-2/IL-R pathway is involved in the regulation of carcinoma cell proliferation. Metabolic labelling followed by immunoprecipitation and Western blot results showed that IL-2 in carcinomas was identical to that in human lymphocytes. However, tumour cells did not secrete IL-2 detectable by immunoassays, although membrane-associated IL-2 was detectable on a proportion of these cells cultured in the absence of exogenous IL-2. Antibodies to IL-2 failed to inhibit proliferation of carcinoma cells, but antibodies specific for the ligand-binding site of the IL-2R were growth inhibitory. Growth of tumour cells was also inhibited by the immunosuppressive drugs, cyclosporin A (CsA), FK506 and rapamycin (RPA), known to interfere with the IL-2 pathway in lymphocytes. To further confirm the role of endogenous IL-2 in the growth of carcinomas, tumour cells were incubated with an IL-2-specific antisense oligonucleotide. The treatment was shown to transiently inhibit IL-2 mRNA and IL-2 protein expression as well as proliferation of tumour cells. Tumour cells treated with IL-2-specific antisense oligonucleotide demonstrated increased apoptosis in comparison to untreated or sense oligonucleotide-treated control cells. The data indicate that in human carcinomas, endogenous IL-2 promotes growth and protects tumour cells from apoptosis.
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16 November 2011
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References
Alileche, A, Plaisance, S, Han, DS, Rubinstein, E, Mingari, C, Bellomo, R, Jasmin, C & Azzarone, B (1993) Human melanoma cell line M14 secretes a functional interleukin-2. Oncogene 8: 1791–1796.
Arzt, E, Stelzer, G, Renner, U, Lange, M, Muller, OA & Stalla, GK (1992) Interleukin-2 and interleukin-2 receptor expression in human corticotrophic adenoma and murine pituitary cell cultures. J Clin Invest 90: 1944–1955.
Capcacioli, S, diPasquale, G, Mini, E, Mazzi, T & Quatrone, A (1993) Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum. biochem. Biochem Bioplys Res Commun 197: 818–825.
Chen, K, Demetris, AJ, VanThiel, DH & Whiteside, TL (1987) Double immunoenzyme staining method for analysis of tissue and blood lymphocyte subsets with monoclonal antibodies. Lab Invest 56: 114–119.
Chomczynski, P & Sacci, N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159.
Ciacci, C, Mahida, YR, Dignass, A, Koizumi, M & Podolsky, DK (1993) Functional interleukin-2 receptors on intestinal epithelial cells. J Clin Invest 92: 527–532.
Damjanovich, S, Bene, L, Matko, J, Alileche, A, Goldman, CK, Sharrow, S & Waldmann, TA (1997) Preassembly of interleukin-2 receptor subunits on resting Kit 225 K6 T cells and their modulating by IL-2, IL-7, and IL-12. A fluorescence resonance energy transfer study. Proc Natl Acad Sci 94: 13134–13139.
Degols, G, Leonetti, JP, Mechti, N & Lebleu, B (1991) Antiproliferative effects of antisense oligonucleotides directed to the RNA of c-myc oncogene. Nucleic Acid Res 19: 945–948.
Giri, JG, Anderson, DM, Kumaki, S, Park, LS, Grabstein, KH & Cosman, D (1995) IL-15, a novel T cell growth factor that shares activities and receptor components with IL-2. J Leuk Biol 57: 763–766.
Grabstein, KH, Eisenman, J, Shanebeck, K, Rauch, C, Srinivasan, S, Fung, V, Beers, C, Richardson, J, Schoenborn, MA & Ahdieh, M and et al (1994) Cloning of a T cell growth factor that interacts with the beta chain of the interleukin-2 receptor. Science 264: 965–968.
Heo, DS, Snyderman, C, Gollin, SM, Pan, S, Walker, E, Deka, RB, Barnes, EL, Johnson, JT, Herberman, RB & Whiteside, TL (1989) Biology, cytogenetics, and sensitivity to immunological effector cells of new head and neck squamous cell carcinoma lines. Cancer Res 49: 5167–5175.
Hesslan, J-MJ, Branellec, AI, Pilatte, Y, Lang, P & Lagrue, G (1991) Differentiation between vascular permeability factor and IL-2 in lymphocyte supernatants from patients with minimal-change nephrotic syndrome. Clin Exp Immunol 86: 157–162.
Kalman, VK & Klimpel, GR (1983) Cyclosporin A inhibits the production of gamma interferon (IFN-γ) but does not inhibit production of virus-induced interferon α/β. Cell Immunol 78: 122–129.
Lin, WC, Yasamura, S, Suminami, Y, Sung, MW, Nagashima, S, Stanson, J & Whiteside, TL (1995) Constitutive production of IL-2 by human carcinoma cells, expression of IL-2 receptor, and tumor cell growth. J Immunol 155: 4805–4816.
McMillan, DN, Kernohan, NM, Flett, ME, Heys, SD, Deehan, DJ, Sewell, HF, Walker, F & Eremin, O (1995) Interleukin-2 receptor expression and interleukin-2 localization in human solid tumor cells in situ and in vitro: evidence for a direct role in the regulation of tumor cell proliferation. Int J Cancer 60: 766–772.
Nagashima, S, Kashii, Y, Reichert, TE, Suminami, Y, Suzuki, T & Whiteside, TL (1997a) Human gastric carcinoma transduced with the IL-2 gene: Increased sensitivity to immune effector cells in vitro and in vivo. Int J Cancer 72: 174–183.
Nagashima, S, Reichert, TE, Kashii, Y, Suminami, Y, Chikamatsu, K & Whiteside, TL (1997b) In vitro characteristics of human squamous cell carcinoma of the head and neck cells engineered to secrete interleukin-2. Cancer Gene Therapy 4: 366–376.
Nakanishi, K, Hirose, S, Yoshimoto, T, Ishizashi, H, Hiroishi, K, Tanaka, T, Kono, T, Miyasaka, M, Taniguchi, T & Higashino, K (1992) Role and regulation of interleukin (IL)-2 receptor alpha and beta chains in IL-2-driven B-cell growth. Proc Natl Acad Sci, USA 89: 3351–3555.
Nourse, J, Firpo, E, Flanagan, WM, Coats, S, Polyak, K, Lee, MH, Massaque, J, Crabtree, GR & Roberts, JM (1994) Interleukin-2-mediated elimination of the p27Kipl cyclin-dependent kinase inhibitor prevented by rapamycin. Nature 372: 570–573.
Plaisance, S, Rubinstein, E, Alileche, A, Sahraoui, Y, Krief, P, Augery-Bourget, Y, Jasmin, C, Suarez, H & Azzarone, B (1992) Expression of the interleukin-2 receptor on human fibroblasts and its biological significance. Int Immunol 4: 739–746.
Plaisance, S, Rubinstein, E, Alileche, A, Benoit, P, Jasmin, C & Azzarone, B (1993) The IL-2 receptor present on human embryonic fibroblasts is functional in the absence of P64/IL-2R gamma chain. Int Immunol 5: 843–848.
Rabinowich, H, Vitolo, D, Altarac, S, Herberman, RB & Whiteside, TL (1992) Role of cytokines in the adoptive immunotherapy of an experimental model of human head and neck cancer by human IL-2-activated natural killer cells. J Immunol 149: 340–349.
Reichert, TE, Nagashima, S, Kashii, Y, Stanson, J, Dou, QP & Whiteside, TL (1998a) Interleukin-2 expression in human carcinoma cell lines and its role in cell cycle progression. Proc AACR 39: 152
Reichert, TE, Watkins, S, Stanson, J, Johnson, TJ & Whiteside, TL (1998b) Endogenous IL-2 in cancer cells: a marker of cellular proliferation. J Histochem Cytochem 46: 603–611.
Rimoldi, D, Salvi, S, Hartmann, F, Schreyer, M, Blum, S, Zografos, L, Plaisance, S, Azzarone, B & Carrel, S (1993) Expression of IL-2 receptors in human melanoma cells. Anticancer Res 13: 555–564.
Sander, B, Andersson, J & Andersson, U (1991) Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev 119: 65–93.
Saneto, RP, Altman, A, Knobler, RL, Johnson, HM & De Vellis, Y (1986) Interleukin-2 mediates the inhibition of oligodendrocyte progenitor cell proliferation in vitro. Proc Natl Acad Sci, USA 83: 9221–9225.
Schreiber, SL (1991) Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science 251: 283–287.
Shimizu, Y, Weidmann, E, Iwatsuki, S, Herberman, RB & Whiteside, TL (1991) Characterization of human autotumor-active T cell clones obtained from tumor-infiltrating lymphocytes in liver metastasis of gastric carcinoma. Cancer Res 51: 6153–6162.
Stein, CA & Cheng, YC (1993) Antisense oligonucleotides as therapeutic agents – is the bullet really magical?. Science 261: 1004–1–12.
Suminami, Y, Kashii, Y, Law, JC, Lin, W-C, Stanson, J, Reichert, TE, Rabinowich, H & Whiteside, TL (1998) Molecular analysis of the IL-2 receptor β chain gene expressed in human tumor cells. Oncogene 16: 1309–1317.
Taniguchi, T & Minami, Y (1993) The IL-2/IL-2 receptor system: a current overview. Cell 73: 5–8.
Terada, N, Lucas, JJ, Szepesi, A, Franklin, RA, Domenico, J & Gelfand, EW (1993) Rapamycin blocks cell cycle progression of activated T cells prior to events characteristic of the middle to late G1 phase of the cycle. J Cell Physiol 154: 7–15.
Waldmann, TA (1991) The interleukin-2 receptor. J Biol Chem 266: 2681–2684.
Weidmann, E, Sacchi, M, Plaisance, S, Heo, DS, Yasamura, S, Lin, WC, Johnson, JT, Herberman, RB, Azzarone, B & Whiteside, TL (1992) Receptors for interleukin-2 on human squamous cell carcinoma cell lines and tumor in situ. Cancer Res 52: 5963–5970.
Whiteside, TL, Sung, M-W, Nagashima, S, Chikamatsu, K, Okada, K & Vujanovic, NL (1998) Human tumor antigen-specific T lymphocytes and IL-2 activated natural killer (A-NK) cells: Comparisons of antitumor effects in vitro and in vivo. Clin Cancer Res 4: 1135–1145.
Yasumura, S, Lin, WC, Weidmann, E, Hebda, P & Whiteside, TL (1994) Expression of interleukin-2 receptors on human carcinoma cell lines and tumor growth inhibition by interleukin-2. Int J Cancer 59: 225–234.
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Reichert, T., Kashii, Y., Stanson, J. et al. The role of endogenous interleukin-2 in proliferation of human carcinoma cell lines. Br J Cancer 81, 822–831 (1999). https://doi.org/10.1038/sj.bjc.6690770
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DOI: https://doi.org/10.1038/sj.bjc.6690770
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