Mouse mammary tumors display Stat3 activation dependent on leukemia inhibitory factor signaling

• Autores: Quaglino, A.; Schere-Levy, C.; Romorini, L.; Meiss, R.P.; Kordon, E.C.
• Año de publicación: 2007
• Idioma: inglés
• Tipo de recurso: artículo
• Fuente: Breast Cancer Res. 2007;9(5)
• Materia: 2 (2 amino 3 methoxyphenyl)chromone; crystal violet; leukemia inhibitory factor; mitogen activated protein kinase 1; mitogen activated protein kinase 3; protein antibody; STAT3 protein; leukemia inhibitory factor; leukemia inhibitory factor receptor alpha; Lif protein, mouse; Lifr protein, mouse; messenger RNA; mitogen activated protein kinase 1; mitogen activated protein kinase 3; STAT3 protein; Stat3 protein, mouse; tyrosine; unclassified drug; animal cell; animal experiment; animal model; article; breast tumor; cell survival; female; human; human cell; immunohistochemistry; mouse; nonhuman; phosphorylation; protein analysis; protein expression; protein function; reverse transcription polymerase chain reaction; Western blotting; animal; Bagg albino mouse; cell culture; experimental neoplasm; fluorescent antibody technique; genetics; immunoprecipitation; metabolism; pathology; signal transduction; Animals; Blotting, Western; Cell Survival; Female; Fluorescent Antibody Technique; Immunoprecipitation; Leukemia Inhibitory Factor; Leukemia Inhibitory Factor Receptor alpha Subunit; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Phosphorylation; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; STAT3 Transcription Factor; Tumor Cells, Cultured; Tyrosine
• Institución: Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales

Introduction: It has been demonstrated that leukemia inhibitory factor (LIF) induces epithelium apoptosis through Stat3 activation during mouse mammary gland involution. In contrast, it has been shown that this transcription factor is commonly activated in breast cancer cells, although what causes this effect remains unknown. Here we have tested the hypothesis that locally produced LIF can be responsible for Stat3 activation in mouse mammary tumors. Methods: The studies were performed in different tumorigenic and non-tumorigenic mammary cells. The expression of LIF and LIF receptor was tested by RT-PCR analysis. In tumors, LIF and Stat3 proteins were analyzed by immunohistochemistry, whereas Stat3 and extracellular signal-regulated kinase (ERK)1/2 expression and phosphorylation were studied by Western blot analysis. A LIF-specific blocking antibody was used to determine whether this cytokine was responsible for Stat3 phosphorylation induced by conditioned medium. Specific pharmacological inhibitors (PD98059 and Stat3ip) that affect ERK1/2 and Stat3 activation were used to study their involvement in LIF-induced effects. To analyze cell survival, assays with crystal violet were performed. Results: High levels of LIF expression and activated Stat3 were found in mammary tumors growing in vivo and in their primary cultures. We found a single mouse mammary tumor cell line, LM3, that showed low levels of activated Stat3. Incidentally, these cells also showed very little expression of LIF receptor. This suggested that autocrine/paracrine LIF would be responsible for Stat3 activation in mouse mammary tumors. This hypothesis was confirmed by the ability of conditioned medium of mammary tumor primary cultures to induce Stat3 phosphorylation, activity that was prevented by pretreatment with LIF-blocking antibody. Besides, we found that LIF increased tumor cell viability. Interestingly, blocking Stat3 activation enhanced this effect in mammary tumor cells. Conclusion: LIF is overexpressed in mouse mammary tumors, where it acts as the main Stat3 activator. Interestingly, the positive LIF effect on tumor cell viability is not dependent on Stat3 activation, which inhibits tumor cell survival as it does in normal mammary epithelium. © 2007 Quaglino et al.; licensee BioMed Central Ltd.
Fil:Quaglino, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Schere-Levy, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Romorini, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil:Kordon, E.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.