The effects of naloxone on human breast cancer progression: in vitro and in vivo studies on MDA.MB231 cells

Abstract

Sabrina Bimonte,1,* Antonio Barbieri,2,* Marco Cascella,1,* Domenica Rea,2,* Giuseppe Palma,2 Vitale Del Vecchio,2 Cira Antonietta Forte,1 Francesco Del Prato,1 Claudio Arra,1 Arturo Cuomo2 1Division of Anesthesia and Pain Medicine, Istituto Nazionale Tumori – IRCCS – “Fondazione G. Pascale”, 2Animal Facility Unit, Department of Research, National Cancer Institute “G. Pascale”, Naples, Italy *These authors contributed equally to this work Background: Naloxone is viewed as a specific competitive opioid antagonist acting at the level of opioid receptors (µ, δ, and κ) with blended agonist-adversary or agonist action. The role of naloxone in tumor cell growth has been poorly studied in human cancer cell lines.Materials and methods: In the present study, we report findings from in vitro and in vivo experiments performed to evaluate the effects of naloxone on human breast cancer cell growth and progression. In vitro assays were conducted on estrogen receptor-negative human breast carcinoma cells, MDA.MB231, treated with naloxone at different concentrations (10–100 µM). In vivo experiments were performed on a mouse model of human triple-negative breast cancer generated by using MDA.MB231 injected subcutaneously in mice. Naloxone was daily intraperitoneally injected in mice at 0.357 mg/kg for 2 weeks and at 0.714 mg/kg for the next 2 weeks. Microvessels formation was detected by fluorescein isothiocyanate-dextran (100 µL) injected into the tail vein of mice and confirmed by immunohistochemistry with CD31 on mice tumor sections.Results: In vitro tests showed that the cell proliferation of MDA.MB231 was inhibited by naloxone in a dose-dependent manner, whereas the cell death was increased. In vivo studies demonstrated that tumors of mice treated with naloxone were significantly smaller than those observed in the control groups, as long as naloxone was administered. Finally, naloxone was not able to impair the microvessel formation in tumors of treated mice.Conclusion: Our data showed, for the first time, that naloxone reduced breast cancer progression without affecting angiogenesis. Keywords: naloxone, breast cancer, proliferation, microvessel formation, angiogenesis

Keywords

• Naloxone
• breast cancer
• proliferation
• microvessel formation
• angiogenesis.