Influence of melatonin on invasive and metastatic properties of MCF-7 human breast cancer cells.
Melatonin, the principal pineal gland hormone, exerts a direct antiproliferative effect on estrogen-responsive MCF-7 cells in culture. The purpose of the current study was to investigate the effects of melatonin on the invasion capacity of MCF-7 cells. In vitro, melatonin at physiological doses (1 nM) reduced (P < 0.001) the invasiveness of tumoral cells measured in Falcon invasion chambers. Subphysiological (0.1 pM) and pharmacological concentrations (10 microM) of melatonin failed to inhibit cell invasion. Melatonin was also able to block 17beta-estradiol-induced invasion (P < 0.001). Pretreatment of MCF-7 cells with 1 nM melatonin increased the response of tumoral cells to the anti-invasive effects of this indolamine. To explore possible mechanisms by which melatonin reduces invasiveness, we measured the attachment of MCF-7 cells to a basement membrane, the chemotactic response of the cells, and their type IV collagenolytic activity. The presence of melatonin (1 nM) in the culture medium significantly reduced the ability of MCF-7 cells to attach to the basement membrane; this effect was enhanced by pretreating the cells with the same indolamine (P < 0.001). Melatonin also counteracts the stimulatory effects of 17beta-estradiol on cell adhesion (P < 0.001). The chemotactic response of MCF-7 cells also decreased in the presence of 1 nM melatonin, and this melatonin-induced reduction of cell migration was more effective on cells that were previously incubated for 5 days with melatonin than it was on nonpretreated cells (P < 0.001). The simultaneous addition of 17beta-estradiol and melatonin resulted in a significantly lower chemotactic response than that of 17beta-estradiol-treated cells (P < 0.001). However, type IV collagenolytic activity was not influenced by melatonin. Our results demonstrate that melatonin reduces the invasiveness of MCF-7 cells, causing a decrease in cell attachment and cell motility, probably by interacting with the estrogen-mediated mechanisms of MCF-7 cell invasiveness. In addition, we also studied the influence of melatonin on the expression of two cell surface adhesion molecules (E-cadherin and beta1 integrin) and an intermediate filament protein (vimentin), the expression of which has been correlated with the relative invasive capacity of human breast cancer cells. The culture of tumor cells in the presence of melatonin (1 nM) increased the membrane staining for E-cadherin and beta1 integrin as well as the number of E-cadherin and beta1 integrin immunoreactive cells (P < 0.01). Neither control MCF-7 cells nor those treated with melatonin stained for vimentin. Preliminary in vivo experiments carried out on ovariectomized athymic nude mice implanted with 17beta-estradiol pellets and inoculated with 5 x 10(6) MCF-7 cells in the inguinal mammary fat pad suggest that melatonin could decrease the tumorigenicity of these tumor cells. However, these results need further confirmation. Taken together, our results suggest that melatonin shifts MCF-7 human breast cancer cells to a lower invasive status by increasing the beta1 integrin subunit and E-cadherin expression and promoting the differentiation of tumor cells. Finally, our study points out the existence of the anti-invasive actions of melatonin as a part of the oncostatic action of melatonin.
Cos
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Department of Physiology and Pharmacology, School of Medicine, University of Cantabria, Santander, Spain.
Fernández
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R
Güézmes
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A
Sánchez-Barceló
E J
EJ
D000273:Adipose Tissue; D000818:Animals; D000970:Antineoplastic Agents; D001485:Basement Membrane; D001943:Breast Neoplasms / Q000473:pathology* / Q000503:physiopathology; D015820:Cadherins / Q000096:biosynthesis; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology*; D017364:Collagenases / Q000378:metabolism; D017077:Culture Media, Conditioned; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006801:Humans; D019012:Integrin beta1 / Q000096:biosynthesis; D008321:Mammary Glands, Animal; D020780:Matrix Metalloproteinase 9; D008550:Melatonin / Q000494:pharmacology* / Q000502:physiology; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D014746:Vimentin / Q000096:biosynthesis
D000970:Antineoplastic Agents; D015820:Cadherins; D017077:Culture Media, Conditioned; D019012:Integrin beta1; D014746:Vimentin; D004958:Estradiol; D017364:Collagenases; D020780:Matrix Metalloproteinase 9; D008550:Melatonin