Neurotensin counteracts apoptosis in breast cancer cells
Section snippets
Materials and methods
Cell culture and synchronization. MCF-7 cells were grown in Dulbecco's modified Eagle's medium (Invitrogen, USA) without phenol red, supplemented with 10% fetal calf serum and 2 mM glutamine. After 24 h of subculture, cells were synchronized with two 16-h pulses of 2 mM thymidine, separated by a 12-h wash in normal media. Cells were treated with 10 nM JMV 449 (H-LysΨ(CH2NH)Lys-Pro-Tyr-Ile-Leu-OH) (Neosystem, France), a potent and stable pseudopeptide NT agonist [16]. The media containing the
NT-1 receptor expression and binding characteristic in MCF-7 cells
Binding studies carried out on MCF-7 cells displayed a specific and saturable binding for NT. Kd and Bmax (Fig. 1A) revealed binding characteristics corresponding to those previously established in other cells for NT-1 receptor [17]. NT-1 receptor mRNA concentration was measured in MCF-7 cells by quantitative RT-PCR (Fig. 1A) [18]. The number of NT-1 receptor mRNA per μg of total RNA in MCF-7 is 10-fold higher than in normal peripheral tissues and absent in normal epithelial breast cells (data
Discussion
During the last decade, neuropeptides and their associated signaling pathways have been increasingly implicated in the development of human cancers [23]. Because of their physiological and cellular effects, a growing interest has focused on NT and its NT-1 receptor. While NT growth effects have been widely demonstrated, the corresponding cellular events are poorly depicted. In this study we demonstrate for the first time, the anti-apoptotic action of NT in the mammary adenocarcinoma cell line,
Acknowledgements
The authors wish to express many thanks to Dr. Neil Insdorf for his precious help in the writing of the manuscript and for helpful discussions. The authors also thank, Dr. S.J. Korsmeyer for the generous gift of Bcl-2 promoter. This work was supported by INSERM and “la ligue nationale contre le cancer,” Grant #75/00-RS/40. Sonia Somaı̈ was supported by fellowships from the Ministère de l'Enseignement Supérieur et de la Recherche and the Fondation pour la Recherche Médicale.
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