[en] BACKGROUND: Neoplasia can results from a lack of cell elimination by apoptosis. In order to determine if mechanisms controlling apoptosis are disturbed during neoplastic transformation in a model of murine radio-induced thymic lymphomas, we have assessed the kinetics of p53, Bax and Bcl-2 in situ expression after induction of thymic apoptosis by irradiation or glucocorticoids at first in normal mice. MATERIALS AND METHODS: TUNEL method was used for in situ detection of apoptosis and protein expression was determined by indirect immunohistochemistry. RESULTS: After hydrocortisone injection, levels of p53 and Bax, but not Bcl-2, expression were raised. A whole body sublethal irradiation led to an increase of p53 and Bcl-2, but not Bax, expression. CONCLUSIONS: This is the first in vivo report of in situ protein expression in the thymus after apoptogenic treatments of mice. The results suggest that Bax could be involved in glucocorticoid-mediated apoptosis. The increased levels of Bcl-2 expression are discussed.
Kaplan HS: On the natural history of the murine leukemias. Cancer Res 27: 1325-1340, 1967.
Boniver J, Decleve A, Lieberman M, Honsik C, Travis M, Kaplan HS: Marrow thymus interactions during radiation leukemogenesis in C57BL/Ka mice. Cancer Res 41: 390-392, 1981.
Haran Ghera N, Rubio N, Leef F, Goldstein G: Characteristics of preleukemia cells induced in mice. Cell Immunol 37: 308-314, 1978.
Defresne MP, Greimers R, Lenaerts P, Boniver J: Effects of marrow grafting on preleukemic cells and thymic nurse cells in C57BL/Ka mice after a leukemogenic split dose irradiation. J Natl Cancer Inst 77: 1079-1085, 1986.
Humblet C, Deman J, Franzen R, Greimers R, Boniver J, Defresne MP: Spontaneous and induced apoptosis after a whole body radiation exposure: experimental approaches. Observations in radio-induced thymic lymphomagenesis. Stem Cells 13 (suppl 1): 129-135, 1995.
Sellins KS and Cohen JJ: Gene induction by gamma irradiation leads to DNA fragmentation in lymphocytes. J Immunol 139: 3199-3206, 1987.
Blomgren H and Andersson B: Characteristics of the immunocon ∼ petent cells in the mouse thymus: cell population changes during cortisone-induced atrophy and subsequent regeneration. Cell Immunol 1: 545-560, 1971.
Cohen JJ and Duke RC: Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J Immunol 132: 38-42, 1984.
Selivanova G and Wiman KG: p53: a cell cycle regulator activated by DNA damage. Adv Cancer Res 66: 143-180, 1995.
Cory S: Regulation of lymphocyte survival by the bcl-2 gene family. Annu Rev Immunol 13: 513-543, 1995.
Yin C, Knudson CM, Korsmeyer SJ, Van Dyke T: Bax suppresses tumorigenensis and stimulates apoptosis in vivo. Nature 385: 82-85, 1995.
Mc Donnell TJ, Deane N, Platt FM, Nunez G, Jaeger U, Mc Kearn JP, Korsmeyer SJ: bcl-2-immunoglobulin transgenic mice demonstrate extended B cell survival and follicular lymphoproliferation.
Hockenbery DM, Zutter M, Hickey W, Nahm M, Korsmeyer SJ: BCL-2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci USA 88: 6961-6965.
Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T: p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362: 847-849, 1993.
Clarke AR, Purdie CA, Harrison DJ, Morris RG, Bird CC, Hooper ML, Wyllie AH: Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 362: 849-852, 1993.
Sentman CL, Shutter JR, Hockenbery D, Kanagawa O, Korsmeyer SJ: bcl-2 inhibits multiple forms of apoptosis but not negative selection in thymocytes. Cell 67: 879-888, 1991.
Strasser A, Harris AW, Cory S: bcl-2 transgene inhibits T cell death and perturbs thymic self-censorsip. Cell 67: 889-899, 1991.
Lassus P, Ferlin M, Piette J, Hibner U: Anti-apoptotic activity of low levels of wild- type p53. EMBO J 15 (17): 4566-4573, 1996.
Surh CD and Sprent J: T-cell apoptosis detected in situ during positive and negative selection in the thymus. Nature 372: 100-103, 1994.
Knudson CM, Tung KSK, Tourtelotte WG, Brown GAJ, Korsmeyer SJ: Bax-deficient mice with lymphoid hyperplasia and male germ cell death. Science 270: 96-99, 1995.
Brady HJM, Salomons GS, Bobeldijk RC, Berns AJM: T cells from box a transgenic mice show accelerated apoptosis in response to stimuli but do not show restored DNA damage-induced cell death in the absence of p53. EMBO J 15 (6): 1221-1230, 1996.
Le PT, Maecker HT, Cook JE: In situ detection and characterization of apoptotic thymocytes in human thymus. Expression of Bcl-2 in vivo does not prevent apoptosis. J Immunol 154: 4371-4378, 1995.
Veis DJ, Sentman CL, Bach EA, Korsmeyer SJ: Expression of the Bcl-2 protein in murine and human thymocytes and in peripheral T lymphocytes. J Immunol 151: 2546-2554, 1993.
Grillot AM, Merino R, Nunez G: Bcl-xL displays restricted distribution during T cell development and inhbits multiple forms of apoptosis but not clonal deletion in transgenic mice. J Exp Med 182: 1973-1983, 1995.
Ma A, Pena JC, Chang B, Margosian E, Davidson L, Alt FW, Thompson CB: Bclx regulates the survival of double-positive thymocytes. Proc Natl Acad Sci USA 92: 4763-4767, 1995.
Motoyama N, Wang F, Roth KA, Sawa H, Nakayama KI, Nakayama K, Negishi I, Senju S, Zhang O, Fujii S, Loh DY: Massive cell death of immature hematopoietic cells and neurons in Bcl-x-deficient mice. Science 267: 1506-1510, 1995.
Chao DT, Linette GP, Boise LH, White LS, Thompson CB, Korsmeyer SJ: Bcl-xL and Bcl-2 repress a common pathway of cell death. J Exp Med 182: 821-828, 1995.
Merville P, Déchanet J, Desmouliere A, Durand I, de Bouteiller O, Garrone P, Banchereau J, Liu YJ: Bcl-2+ tonsillar plasma cells are rescued from apoptosis by bone marrow fibroblasts. J Exp Med 183: 227-236, 1996.
Hugo P, Kappler JW, Godfrey DI, Marrack PC: A cell line that can induce thymocyte positive selection. Nature 360: 679-682, 1992.
Hugo P, Kappler JW, McCormack JE, Marrack P: Fibroblasts can induce thymocyte positive selection in vivo. Proc Natl Acad Sci USA 90: 10335-10339, 1993.
Tsujimoto Y, Croce CM: Analysis of the structure, transcripts and protein products of bcl-2, the gene involved in human follicular lymphoma. Proc Natl Acad Sci USA 83: 5214-5218, 1986.
Tanaka S, Saito K, Reed JC: Structure-function analysis of the bcl-2 oncoprotein. J Biol Chem 268: 10920-10926, 1993.
Cheng EHY, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K, Hardwich JM: Conversion of Bcl-2 to a Bax-like death effector by caspases. Science 278: 1966-1968, 1997.
Krajewski S, Tanaka S, Takayama S, Schibler MJ, Fenton W, Reed JC: Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum and outer mitochondrial membranes. Cancer Res 53: 4701-4714.