RESUMO
Exposure of male hamsters to short daily photoperiods (1 hour of light and 23 hours of darkness daily for 9 weeks led to total involution of the testes and accessory sex organs (seminal vesicles and coagulating glands). Pituitary levels of immunoreactive prolaction also decreased by about 60 percent after dark exposure. The inhibitory effects of darkness on the reproductive organs were prevented either by pinealectomy or by the subcutaneous implantation of a melatonin-beeswax pellet into the animals each week. Both pinealectomy and melatonin treatment also returned pituitary levels of prolactin toward normal. The results suggest that melatonin is not the pineal antigonadotrophic factor in the male golden hamster.
Assuntos
Gonadotropinas Hipofisárias/metabolismo , Glândula Pineal/fisiologia , Animais , Peso Corporal , Ritmo Circadiano , Cricetinae , Escuridão , Luz , Masculino , Melatonina/farmacologia , Melatonina/fisiologia , Tamanho do Órgão , Glândula Pineal/efeitos dos fármacos , Adeno-Hipófise/análise , Prolactina/análise , Prolactina/sangue , Prolactina/metabolismo , Próstata/anatomia & histologia , Próstata/efeitos dos fármacos , Radioimunoensaio , Glândulas Seminais/anatomia & histologia , Glândulas Seminais/efeitos dos fármacos , Especificidade da Espécie , Testículo/anatomia & histologia , Testículo/efeitos dos fármacosRESUMO
Since melatonin, the major hormone of the pineal gland, has been shown to inhibit the growth of mammary tumors in animal models of human breast cancer, we examined the hypothesis that this indoleamine has the potential to inhibit breast cancer growth by directly inhibiting cell proliferation as exemplified by the growth of the estrogen-responsive human breast cancer cell line MCF-7 in culture. Concentrations of melatonin (10(-9) M; 10(-11) M), corresponding to the physiological levels present in human blood during the evening hours, significantly inhibited (P less than 0.001) cell proliferation by as much as 60% to 78% as measured by either DNA content or hemocytometer cell counts. Melatonin's inhibitory effect was reversible since the logarithmic growth of MCF-7 cells was restored after melatonin-containing medium was replaced with fresh medium lacking melatonin. Not only was the inhibitory effect of melatonin absent at either pharmacological (10(-7) M; 10(-5) M) or subphysiological (10(-15) M; 10(-13) M) concentrations, but melatonin also failed to inhibit the proliferation of either human foreskin fibroblasts or the estrogen receptor-positive human endometrial cancer cell line RL95-2. Both transmission and scanning electron microscopy revealed several morphological changes that correlated with melatonin's inhibition of cell growth. After just 4 days of exposure to melatonin, MCF-7 cells exhibited reduced numbers of surface microvilli, nuclear swelling, cytoplasmic and ribosomal shedding, disruption of mitochondrial cristae, vesiculation of the smooth endoplasmic reticulum, and an increase in the numbers of autophagic vacuoles. These results support the hypothesis that melatonin, at physiological concentrations, exerts a direct but reversible, antiproliferative effect on MCF-7 cell growth in culture. This antiproliferative effect is associated with striking changes in the ultrastructural features of these cells suggestive of a sublethal but reversible cellular injury.
Assuntos
Neoplasias da Mama/patologia , Melatonina/farmacologia , Neoplasias da Mama/ultraestrutura , Divisão Celular/efeitos dos fármacos , DNA de Neoplasias/análise , Feminino , Humanos , Células Tumorais Cultivadas , Vacúolos/efeitos dos fármacosRESUMO
Dietary intake of the n-6 fatty acid (FA) linoleic acid (LA) has a strong growth-promoting effect on many rodent tumors and human tumor xenografts grown in immunodeficient rodents. n-3 FAs such as alpha-linolenic and eicosapentaenoic acids (EPAs), which differ from LA and arachidonic acid, respectively, by only a single double bond in the n-3 position, are recognized cancer chemopreventive and anticachectic agents. Understanding how this seemingly small structural difference leads to such remarkable functional differences has been a challenge. In a previous study, we showed that LA uptake, [3H]thymidine incorporation into DNA, and total DNA content were decreased in tissue-isolated hepatoma 7288CTC perfused in situ with arterial blood containing alpha-linolenic acid, EPA, or docosahexaenoic acids. The Ki for the inhibition of LA uptake and [3H]thymidine incorporation by alpha-linolenic acid was 0.18 and 0.25 mM, respectively. Here we show that the addition of alpha-linolenic acid or EPA to arterial blood inhibits tumor FA uptake, including LA, and the subsequent conversion of LA to the mitogen 13-hydroxyoctadecadienoic acid (13-HODE) in vivo and during perfusion in situ. [3H]Thymidine incorporation during perfusion in situ was also inhibited. Addition of 13-HODE to the arterial blood reversed the inhibition of [3H]thymidine incorporation but had no effect on FA uptake. These two n-3 FAs also inhibited FA transport in inguinal fat pads in vivo and during perfusion in situ in fed (FA uptake) and fasted (FA release) rats. The effects of EPA and talinolenic acid on transport of saturated, monounsaturated, and n-6 polyunsaturated FAs in hepatoma 7288CTC and inguinal fat pads during perfusion in situ were reversed by the addition of forskolin (1 microM), pertussis toxin (0.5 microg/ml), or 8-bromo-cyclic AMP (10 microM) to the arterial blood. We conclude that the antitumor and anticachectic effects of n-3 FAs on hepatoma 7288CTC and inguinal fat pads in vivo result from an inhibition of FA transport. These inhibitions are mediated by a putative n-3 FA receptor via a Gi protein-coupled signal transduction pathway that decreases intracellular cyclic AMP. A specific decrease in LA uptake and its conversion to the mitogen 13-HODE causes the tumor growth inhibition.
Assuntos
Antineoplásicos/farmacologia , Caquexia/tratamento farmacológico , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos/farmacocinética , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Ácido alfa-Linolênico/farmacologia , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Caquexia/sangue , Colforsina/farmacologia , AMP Cíclico/metabolismo , Ácidos Graxos/antagonistas & inibidores , Ácidos Graxos/metabolismo , Proteínas de Ligação ao GTP/metabolismo , Ácidos Linoleicos/biossíntese , Neoplasias Hepáticas Experimentais/sangue , Neoplasias Hepáticas Experimentais/metabolismo , Masculino , Transplante de Neoplasias , Ratos , Ratos Endogâmicos BUF , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Timidina/metabolismoRESUMO
Melatonin, the chief hormone secreted by the pineal gland, has been previously shown to inhibit human breast cancer cell growth at the physiological concentration of 1 nM in vitro. In this study, using the estrogen receptor (ER)-positive human breast tumor cell line MCF-7, we have shown that 10 microM L-buthionine-[S,R]-sulfoximine (L-BSO), an inhibitor of gamma-glutamylcysteine synthetase (the rate-limiting enzyme in glutathione synthesis), blocks the oncostatic action of 1 nM melatonin over a 5-day incubation, indicating that glutathione is required for melatonin action. The result was repeated with ZR75-1 cells, suggesting that the glutathione requirement is a general phenomenon among ER+ breast cancer cells. Addition of exogenous glutathione (1 microM) to L-BSO-treated groups restored the melatonin response in both cell lines. Further demonstration of the importance of glutathione was shown using the ER- breast tumor cell line HS578T, which is normally unresponsive to melatonin. Growth in this cell line was inhibited in the presence of 1 microM ethacrynic acid (an inhibitor of glutathione S-transferase) plus 1 nM melatonin, and this effect was blocked with 10 microM L-BSO. We also observed a steady decrease of intracellular glutathione in MCF-7 cells over a 5-day incubation, suggesting that these cells metabolize glutathione differently than do normal cells.
Assuntos
Neoplasias da Mama/patologia , Glutationa/fisiologia , Melatonina/farmacologia , Antimetabólitos Antineoplásicos/farmacologia , Antineoplásicos Hormonais/farmacologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Butionina Sulfoximina/farmacologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Diuréticos/farmacologia , Ácido Etacrínico/farmacologia , Feminino , Glutationa/metabolismo , Glutationa/farmacologia , Humanos , Melatonina/fisiologia , Receptores de Estrogênio/fisiologia , Tamoxifeno/farmacologia , Células Tumorais CultivadasRESUMO
Growth of hepatoma 7288CTC in male Buffalo rats is directly dependent on uptake of linoleic acid (LA) from the arterial blood. One to 5% of the LA taken up is converted to 13-hydroxyoctadecadienoic acid (HODE), an agent that enhances epidermal growth factor-dependent mitogenesis. The role of 13-HODE in LA-dependent growth of solid tumors is not known. In this study, we examined LA uptake and 13-HODE formation on growth of tissue-isolated hepatoma 7288CTC in vivo and on [3H]thymidine incorporation and DNA content during perfusion in situ. Fatty acid uptake and metabolite release were determined from arteriovenous difference measurements. Tumor-bearing and blood donor rats were fed either LA-sufficient or -deficient diets. Hepatoma 7288CTC removed LA from the arterial blood and released 13-HODE [and a small amount of 13-ketooctadecadienoic acid (KODE)] into the venous blood both in vivo and during perfusion. Treatment with the lipoxygenase inhibitor nordihydroguaiaretic acid (10 microM) did not affect tumor LA uptake, but inhibited release of 13-HODE and 13-KODE in vivo and during perfusion, suppressed growth in vivo, and inhibited [3H]thymidine incorporation during perfusion. The addition of 13-HODE to the nordihydroguaiaretic acid-containing whole blood perfusate increased the rate of [3H]thymidine incorporation 10 times and nearly doubled tumor DNA content; the addition of 13-KODE or 9-HODE had no effect. 13-HODE and 13-KODE were not released from tumors growing in rats fed a LA-deficient diet, and the rates of tumor growth in vivo and [3H]thymidine incorporation during perfusion were decreased. The addition of 13-HODE to the LA-deficient blood perfusate promoted tumor 13-HODE uptake and a dose-dependent increase in [3H]thymidine incorporation and tumor DNA content. These results provide strong evidence that 13-HODE is the mitogenic signal responsible for LA-dependent growth in hepatoma 7288CTC in vivo.
Assuntos
Ácido Linoleico/farmacologia , Ácidos Linoleicos/farmacologia , Neoplasias Hepáticas Experimentais/patologia , Animais , Transporte Biológico , Divisão Celular/efeitos dos fármacos , DNA de Neoplasias/biossíntese , Ácido Linoleico/sangue , Ácido Linoleico/metabolismo , Ácidos Linoleicos/sangue , Ácidos Linoleicos/metabolismo , Neoplasias Hepáticas Experimentais/metabolismo , Masculino , Masoprocol/farmacologia , Ratos , Ratos Endogâmicos BUF , Timidina/metabolismoRESUMO
The growth of rat hepatoma 7288CTC in vivo is stimulated by the uptake of linoleic acid (LA) and its metabolism to 13-hydroxyoctadecadienoic acid (13-HODE), an important mitogenic signaling molecule within this tumor. Conversely, the growth of a variety of experimental cancers in vivo is inhibited by either physiological or pharmacological levels of the pineal gland hormone melatonin, although the mechanism(s) are unknown. We tested the hypothesis that the mechanism of melatonin's anticancer action in vivo involves the inhibition of tumor LA uptake and metabolism to 13-HODE in hepatoma 7288CTC. Tumor uptake of LA and release of 13-HODE, measured in tissue-isolated rat hepatoma 7288CTC at 4-h intervals over a 24-h period, were highest during the light phase and lowest during the mid-dark phase, when plasma melatonin levels were lowest and highest, respectively. Pinealectomy eliminated this rhythm of tumor LA uptake and 13-HODE production, indicating that it was driven by the circadian melatonin rhythm. Perfusion of tissue-isolated tumors in situ with melatonin (1 nM) rapidly and reversibly inhibited the uptake of plasma fatty acids (FAs), including LA, and its metabolism to 13-HODE. These inhibitory effects of melatonin on tumor FA uptake and 13-HODE release were completely reversed by perfusion of tumors in situ with melatonin receptor antagonist S-20928, pertussis toxin, forskolin, or 8-bromo-cAMP. Perfusion of tumors in situ with melatonin also decreased tumor [3H]thymidine incorporation and DNA content; these effects on DNA synthesis were also prevented by the coperfusion of tumors with melatonin and S-20928, pertussis toxin, forskolin, 8-Br-cAMP, or 13-HODE. Pinealectomy stimulated tumor growth, LA uptake and metabolism to 13-HODE, and FA storage in hepatoma 7288CTC, whereas melatonin administration (200 microg/day) was inhibitory in vivo. Northern blot analysis revealed that, compared with normal liver tissue, hepatoma 7288CTC overexpressed mRNA transcripts for a plasma membrane-associated FA transport protein (FATP). FATP mRNA expression was unaffected by the treatment of tumor-bearing rats with daily afternoon melatonin injections or exposure to constant light. These results support a novel mechanism of tumor growth inhibition by melatonin involving a melatonin receptor-mediated suppression of cAMP levels, resulting in diminished tumor FA transport, possibly via decreased FATP function. The inhibition of these signal transduction events by melatonin culminates in the suppression of LA uptake, LA metabolism to the mitogenic signaling molecule 13-HODE, and cancer growth.
Assuntos
Ácido Linoleico/metabolismo , Ácidos Linoleicos/metabolismo , Neoplasias Hepáticas Experimentais/patologia , Neoplasias Hepáticas Experimentais/fisiopatologia , Melatonina/farmacologia , Receptores de Superfície Celular/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , Transdução de Sinais/fisiologia , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Ritmo Circadiano , Colforsina/farmacologia , Ácidos Linoleicos/farmacologia , Masculino , Modelos Biológicos , Naftalenos/farmacologia , Perfusão , Toxina Pertussis , Ratos , Ratos Endogâmicos BUF , Receptores de Superfície Celular/antagonistas & inibidores , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Receptores de Melatonina , Transdução de Sinais/efeitos dos fármacos , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Because the pineal hormone melatonin has been implicated in affecting adiposity in rats and fatty acid transport in certain rat tumor models, we tested whether melatonin regulates lipolysis in a normal cell system in vitro. Adipocytes were isolated from the inguinal fat pads (i.e. sc fat) of Sprague Dawley male rats during mid-light phase. Lipolysis was stimulated with isoproterenol (3 microM), and cells were incubated for 4 h in the presence or absence of a physiological circulating concentration of melatonin (1 nM). Lipolysis was measured by determining the amount of glycerol present in the incubation buffer, expressed as nmol glycerol/mg cellular fatty acid. We observed a 20- to 30-fold stimulation of basal lipolysis by isoproterenol, and this stimulation was inhibited 50--70% by melatonin. Melatonin exhibited this effect over a wide range of concentrations tested (100 pM-1 microM) with an IC(50) of approximately 500 pM. The effect by melatonin (1 nM) was completely blocked by pertussis toxin (50 ng/ml), by 8-bromo-cAMP (10 nM), and by the melatonin receptor antagonist S-20928 (1 nM). These results suggest that the antilipolytic effect occurs through one of the G(i) protein-coupled melatonin receptors because we have shown that both the mt(1) (Mel 1a) and MT(2) (Mel 1b) melatonin receptors are expressed in inguinal adipocytes. Melatonin inhibition of lipolysis was not observed in adipocytes isolated from rat epididymal fat pads (i.e. visceral fat), even though these cells also express both the mt(1) and MT(2) receptors. The results indicate that physiological circulating concentrations of melatonin inhibit isoproterenol-induced lipolysis in rat adipocytes via a G protein-coupled melatonin receptor-mediated signal transduction pathway in a site-specific manner.
Assuntos
Adipócitos/metabolismo , Tecido Adiposo/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Isoproterenol/farmacologia , Lipólise/fisiologia , Melatonina/fisiologia , Receptores de Superfície Celular/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Adipócitos/efeitos dos fármacos , Tecido Adiposo/citologia , Tecido Adiposo/efeitos dos fármacos , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Epididimo , Virilha , Lipólise/efeitos dos fármacos , Masculino , Melatonina/antagonistas & inibidores , Melatonina/farmacologia , Naftalenos/farmacologia , Concentração Osmolar , Toxina Pertussis , Ratos , Ratos Sprague-Dawley , Receptores de Superfície Celular/antagonistas & inibidores , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Receptores de Melatonina , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Estrogen-progesterone (EP) treated adult male rats were injected intravenously (iv) with 0.1, 1 or 10 mug arginine vasotocin (AVT) or Ringers lactate solution. A significant dose-related rise in plasma prolactin was evident 10 min after injection with AVT. In a second experiment, sham-operated or pinealectomized EP-treated male rats were injected iv with 0.1 or 1 mug AVT or diluent. Plasma prolactin was significantly elevated in both sham-operated and pinealectomized groups at both 10 and 20 min post-injection of 1 mug AVT. These results indicate that AVT has prolactin-releasing activity and that this activity is not dependent upon the presence of an intact pineal gland.
Assuntos
Glândula Pineal/fisiologia , Prolactina/metabolismo , Vasotocina/farmacologia , Animais , Relação Dose-Resposta a Droga , Estradiol/farmacologia , Masculino , Progesterona/farmacologia , Ratos , Estimulação QuímicaRESUMO
A sc injection of 2 micrograms arginine vasotocin (AVT) administered every 2 h for a 25-h period starting 1 h after castration of adult male rats caused a 43% reduction (P less than 0.05) in plasma levels of LH compared to diluent-treated castrated controls. In Exp 2, a sc injection of 5 micrograms AVT or diluent was administered to intact or acutely castrated male rats every 3 h for 48 h. After AVT administration, both plasma LH (P less than 0.001) and FSH (P less than 0.05) were significantly reduced, with a concomitant increase in pituitary levels of these gonadotropins. Using the same injection regimen in Exp 3, 250 micrograms melatonin had no effect on plasma or pituitary levels of LH in castrated male rats. In the fourth experiment, neither 1 IU arginine vasopressin nor 1 IU oxytocin had an effect on plasma or pituitary levels of LH, whereas 1 IU of AVT significantly lowered plasma levels of LH (P less than 0.05) and prevented the fall in the pituitary level of this gonadotropin (P less than 0.01) when compared to diluent-treated castrated control rats. Oxytocin (1 IU) significantly inhibited (P less than 0.01) plasma levels of FSH and raised plasma levels of PRL (P less than 0.01) in castrated male rats. No effect on plasma titers of PRL were observed after treatment of castrated rats with AVT in Exp 2 or 4. It is concluded that in acutely castrated male rats, AVT could possibly act either on the hypothalamus and/or the pituitary to affect pituitary and plasma levels of gonadotropins.
Assuntos
Hormônio Foliculoestimulante/metabolismo , Hormônio Luteinizante/metabolismo , Melatonina/farmacologia , Hipófise/metabolismo , Prolactina/metabolismo , Vasotocina/farmacologia , Animais , Castração , Hormônio Foliculoestimulante/sangue , Hormônio Luteinizante/sangue , Masculino , Hipófise/efeitos dos fármacos , Prolactina/sangue , RatosRESUMO
The effects of crude extracts of bovine, rat, and human pineal glands on prolactin (PRL) release were studied using an in vitro system. In addition, the effects of a known pineal constituent, arginine vasotocin (AVT), and crude bovine pineal extract (bPE) on PRL secretion were studied in vivo. Normal male rat hemipituitaries (HP), incubated with bPE (13 mg tissue/HP)released 200%, 150%, and 285% more PRL into the medium than did their corresponding untreated control halves incubated in either Medium 199 alone, hypothalamic extract, or cerebral cortical extract, respectively. HP incubated with either rat (6 mg of tissue/HP) or human (25 mg of tissue/HP) pineal extract released 110% and 75% more PRL, respectively, than did their corresponding untreated control halves. HP exposed to 10 mg tissue eq of either bovine pineal fraction A1 or bovine pineal fraction A3 released 88% and 63%, respectively, less PRL than did their corresponding untreated control halves incubated in Krebs-Ringer Bicarbonate (KRB) medium. Quantitites of melatonin, thyrotropin-releasing hormone (TRH), or estrogen, comparable to those found in the pineal, had no significant effect on PRL secretion in vitro. The iv injection of either bPE (90 mg tissue/rat) or AVT (10 mug/rat) into estrogen and progesterone-treated male rats resulted in a 40% and 138% increase, respectively, in plasma PRL titers, 10 min after injection, over pre-injection control levels. The per cent of increase in plasma PRL levels in these animals was significantly greater than that observed in control rats receiving either saline or cortical extract. The results suggest that crude extracts of pineal glands of three different species contain prolactin-releasing factor (PRF) activity which is probably not due to any endogenous melatonin, TRH, or estrogen that may be present. Conversely, two bovine pineal fractions, A1 and A3, appeared to exhibit prolactin-inhibiting factor (PIF) activity. We have concluded that the pineal gland may serve as an alternate or supplemental source of PRF and/or PIF.
Assuntos
Glândula Pineal , Prolactina/metabolismo , Extratos de Tecidos/farmacologia , Idoso , Animais , Bovinos , Córtex Cerebral , Estradiol/farmacologia , Feminino , Humanos , Técnicas In Vitro , Masculino , Melatonina/farmacologia , Pessoa de Meia-Idade , Glândula Pineal/análise , Glândula Pineal/metabolismo , Fatores Inibidores da Liberação da Prolactina/análise , Ratos , Estimulação Química , Hormônio Liberador de Tireotropina/análise , Hormônio Liberador de Tireotropina/farmacologia , Vasotocina/farmacologiaRESUMO
Maintaining adult male golden hamsters in short daily photoperiods (1hr of light and 23hr of darkness daily; LD 1.23) for 12 weeks caused the tests and accessory sex organs to atrophy and also led to significant depressions in pituitary LH and prolactin levels. If hamsters that were kept in LD 1.23 CYCLES RECEIVED WEEKLy subcutanceous implants of either a melanioninbeeswax or a 5-methoxtrypotophol-beeswax pellet (1 mgindole in 24 mg beeswax) the testes and accessory sex organs failed to involute and pituitary LH levels did not drop. Both treatments also retarded the depression in hypophyseal prolactin retarded the depression in hypophyseal prolactin levels. Treatment (twice daily on weekdays and once daily on weekends) with 1.5 mug injections of synthetic LRH (in 0.2 ml 8% gelatin) did not prevent gonadal or accessory organ atrophy while it further depressed hypophyseal LRH treatment, plasma LH titers were significantly elevated. The reproductive organs of hamsters that were moved from short (LD1;I 23) to long (LD 14;10) daily photoperiods regenerated within 8 weeks. This light-induced restoration of the gonads was not prevented or retarded by the weekly implantation of either melantonin-beeswax or 5-methoxytrptophol-beeswax pellets. The results suggest that in the male golden hamster neither melatonin nor 5-methoxtryptophol accounts for the antigonadotrophic activity of the pineal gland.
Assuntos
Indóis/farmacologia , Hormônio Luteinizante/metabolismo , Melatonina/farmacologia , Glândula Pineal/fisiologia , Reprodução/efeitos dos fármacos , Testículo/fisiologia , Animais , Peso Corporal , Cricetinae , Escuridão , Hormônio Liberador de Gonadotropina/farmacologia , Luz , Masculino , Éteres Metílicos/farmacologia , Tamanho do Órgão , Glândula Pineal/efeitos dos fármacos , Adeno-Hipófise/metabolismo , Prolactina/metabolismo , Regeneração , Glândulas Seminais/fisiologia , Fatores de TempoRESUMO
Cultured MCF-7 human breast cancer cells were pre-exposed to either melatonin (232 ng/mL) or vehicle for 24 hrs prior to being washed and then re-exposed to either ethanol-vehicle or varying concentrations of tamoxifen (37.1 ng/mL, 3.71 micrograms/mL, 371 micrograms/mL) or melatonin (2.32 pg/mL, 232 ng/mL, 23.2 ng/mL) for 5 additional days. Only 371 ng/mL tamoxifen caused a 38% growth inhibition of cells pre-exposed to vehicle whereas all concentrations of tamoxifen inhibited the growth of melatonin pre-exposed cells by 28% to 61% in a dose-dependent manner. Melatonin pre-exposure, potentiated the inhibitory effect of only 232 ng/mL melatonin. Comparison of IC50 values indicate that tamoxifen is approximately a 100 times more potent inhibitor of breast cancer cell growth following the pretreatment of cells with a physiological concentration of melatonin. These results indicate that melatonin has the capability to augment the inhibitory actions of tamoxifen, and to a lesser extent itself, on human breast cancer cell growth.
Assuntos
Neoplasias da Mama/patologia , Melatonina/farmacologia , Tamoxifeno/farmacologia , Divisão Celular/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Células Tumorais CultivadasRESUMO
Only physiological levels of melatonin exert an antiproliferative effect on MCF-7 breast cancer cells grown in an anchorage-dependent culture system. We investigated melatonin's effect on the anchorage-independent growth of MCF-7 cells as well as the dose-response characteristics of this indoleamine under clonogenic culture conditions. Melatonin's inhibitory effect, with respect to the number and size of colonies formed, exhibit a linear dose-response curve with pharmacological concentrations producing a maximal inhibition while subphysiological levels of melatonin induce minimal inhibition. These results indicate that cellular attachment may modify the sensitivity of MCF-7 cells towards melatonin.
Assuntos
Antineoplásicos , Neoplasias da Mama/patologia , Melatonina/farmacologia , Glândula Pineal/metabolismo , Adesão Celular , Divisão Celular/efeitos dos fármacos , Meios de Cultura , Depressão Química , Relação Dose-Resposta a Droga , Feminino , Humanos , Tamoxifeno/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/patologia , Ensaio Tumoral de Célula-TroncoRESUMO
The pineal hormone, melatonin, was examined for its capacity to modulate the proliferation of a panel of human breast cancer cell lines. Melatonin inhibited, to a varying extent, the proliferation of all three estrogen-responsive cell lines, but had no effect on estrogen-insensitive breast tumor cell lines. Melatonin was also able to specifically block estrogen-induced proliferation in MCF-7 breast cancer cells. However, this action was abolished in the presence of tamoxifen. Therefore, it appears that the antiproliferative effects of melatonin are mediated through the estrogen-response pathway.
Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Antagonistas de Estrogênios/farmacologia , Estrogênios/fisiologia , Melatonina/farmacologia , Neoplasias Hormônio-Dependentes/tratamento farmacológico , Neoplasias da Mama/patologia , Divisão Celular/efeitos dos fármacos , Interações Medicamentosas , Humanos , Neoplasias Hormônio-Dependentes/patologia , Receptores de Estrogênio/fisiologia , Tamoxifeno/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Tumor linoleic acid uptake and metabolism, and growth are suppressed by melatonin, the synthesis of which is inhibited by light. Linoleic acid, via its mitogenic metabolite 13-hydroxyoctadecadienoic acid (13-HODE) is an important growth stimulant of rat hepatoma 7288CTC. Here we compared the effects of an alternating light:dark cycle (12L:12D), dim light (0.25 lux) present during the dark phase of a diurnal light cycle, and constant light on growth and fatty acid metabolism in hepatoma 7288CTC. Our results show that dim light suppressed melatonin release by the pineal gland, increased tumor linoleic acid uptake and 13-HODE production, and promoted tumor growth as effectively as did constant light.
Assuntos
Escuridão , Luz , Ácido Linoleico/farmacocinética , Animais , Divisão Celular , Progressão da Doença , Ácido Linoleico/metabolismo , Ácidos Linoleicos/metabolismo , Masculino , Melatonina/antagonistas & inibidores , Melatonina/biossíntese , Ratos , Ratos Endogâmicos BUFRESUMO
In experimental trials using the N-nitroso-N-methylurea (NMU)-induced rat mammary tumor model, a significant decrease in tumor incidence (to 5%) was observed in rats treated with melatonin and 9-cis-retinoic acid (9 cRA) compared to controls (55%). Although 9cRA alone decreased tumor incidence to 26%, this response did not reach statistical significance. Tumor incidence was significantly inhibited to 20% in the animals that received melatonin and 9cRA on alternating days. Latency to tumor onset was prolonged in animals receiving either of the combination treatments compared with controls, and tumor multiplicity was also significantly decreased.
Assuntos
Adenocarcinoma/prevenção & controle , Anticarcinógenos/farmacologia , Neoplasias Mamárias Experimentais/prevenção & controle , Melatonina/farmacologia , Tretinoína/farmacologia , Adenocarcinoma/induzido quimicamente , Adenocarcinoma/patologia , Alitretinoína , Animais , Antioxidantes/farmacologia , Peso Corporal/efeitos dos fármacos , Carcinógenos/antagonistas & inibidores , Sinergismo Farmacológico , Quimioterapia Combinada , Estradiol/sangue , Receptor alfa de Estrogênio , Feminino , Sequestradores de Radicais Livres/farmacologia , Neoplasias Mamárias Experimentais/induzido quimicamente , Neoplasias Mamárias Experimentais/patologia , Metilnitrosoureia , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Estrogênio/biossíntese , Receptores do Ácido Retinoico/biossíntese , Receptor alfa de Ácido Retinoico , Útero/anatomia & histologia , Útero/efeitos dos fármacosRESUMO
Many nutritional, hormonal, and environmental factors affect carcinogenesis and growth of established tumors in rodents. In some cases, these factors may either enhance or attenuate the neoplastic process. Recent experiments performed in our laboratory using tissue-isolated rat hepatoma 7288CTC in vivo or during perfusion in situ have demonstrated new interactions among four of these factors. Two agents, dietary linoleic acid (C18:2n6) and "light at night," enhanced tumor growth, and two others, melatonin and n3 fatty acids, attenuated growth. Linoleic acid stimulated tumor growth because it is converted by hepatoma 7288CTC to the mitogen, 13-hydroxyoctadecadienoic acid (13-HODE). Melatonin, the neurohormone synthesized and secreted at night by the pineal gland, and dietary n3 fatty acids are potent antitumor agents. Both inhibited tumor linoleic acid uptake and 13-HODE formation. Artificial light, specifically "light at night," increased tumor growth because it suppressed melatonin synthesis and enhanced 13-HODE formation. Melatonin and n3 fatty acids acted via similar or identical G(i) protein-coupled signal transduction pathways, except that melatonin receptors and putative n3 fatty acid receptors were used. The results link the four factors in a common mechanism and provide new insights into the roles of dietary n6 and n3 polyunsaturated fatty acid intake, "light at night," and melatonin in cancer prevention in humans.
Assuntos
Antioxidantes/uso terapêutico , Ácidos Graxos Insaturados/uso terapêutico , Melatonina/uso terapêutico , Neoplasias/prevenção & controle , Animais , Humanos , Ácido Linoleico/uso terapêuticoRESUMO
Intravenous (iv) administration of 5 microgram of arginine vasotocin (AVT) into urethane-anesthetized, castrated male rats had no effect on plasma prolactin titers as compared to the rise in prolactin levels observed in intact AVT-treated rats. However, when castrated rats were first treated for two days with 2.5 mg/day of testosterone propionate and then challenged with a 5-microgram dose of AVT, the prolactin surge values obtained were comparable to those seen in intact AVT-treated rats. Conversely, treatment of intact rats for two days with 25 mg/day of the anti-androgen, cyproterone acetate, blocked the prolactin-releasing activity of AVT. In a separate experiment, treatment of castrated rats for two days with 2.5 mg/day of testosterone, androsterone or 50 microgram of estradiol benzoate and 25 mg progesterone, completely restored the prolactin-releasing activity of AVT. Similar treatment with 2.5 mg/day of androstenedione or dihydrotestosterone was without effect in restoring this response to AVT. It is concluded that the presence of gonadal steroids is essential to the action of AVT in provoking the release of prolactin in urethane-anesthetized male rats.
Assuntos
Androgênios/farmacologia , Ciproterona/farmacologia , Estradiol/farmacologia , Progesterona/farmacologia , Prolactina/metabolismo , Vasotocina/farmacologia , Androstenodiona/farmacologia , Androsterona/farmacologia , Animais , Castração , Di-Hidrotestosterona/farmacologia , Masculino , Ratos , Testosterona/farmacologiaRESUMO
Treatment of unanesthetized castrated adult male rats every 3 h for 48 h with either 5 microgram of arginine vasotocin (AVT) and/or 1 microgram luteinizing hormone-releasing hormone (LRH) caused a significant inhibition of plasma levels of luteinizing hormone (LH) and compared to castrated control rats receiving diluent only. However, the intravenous (iv) injection of 1 microgram of AVT into urethane-anesthetized male rats which had been castrated for 0, 24 or 48 h did not affect plasma levels of LH at 10, 20 or 60 min following injection compared to their respective diluent-treated castrated control rats. Similarly, the iv injection of either 100 ng, 1 microgram or 10 microgram AVT was unable to acutely affect plasma levels of LH in intact male rats. Following the iv injection of 2 doses of 50 ng LRH spaced 1 h apart in anesthetized castrated male rats, 2 peaks of equal magnitude in plasma LH were noted. Castrated rats treated with 2 injections spaced 1 h apart of LRH + AVT had significantly higher plasma levels of LH than did rats treated with LRH alone. In subsequent studies, both AVT and arginine vasopressin were observed to augment the plasma response of LH to an injection of LRH whereas oxytocin had no effect. A single injection of AVT + LRH significantly augmented the plasma titers of LH compared to levels observed in LRH-treated control rats as did a second injection 1 h later. The administration of cyproterone acetate sc for 2 days by itself had no effect on plasma LH but in conjunction with LRH caused a marked rise in plasma LH compared to intact rats treated with LRH alone. AVT in combination with LRH and cyproterone acetate caused a significant elevation in plasma LH at 60 min post-injection when compared to plasma levels of rats treated with LRH alone or the combination of LRH and cyproterone acetate. It is concluded that acute intravenous injections of AVT augment the LH-releasing activity of LRH; chronic treatment for 48 h, however, with LRH + AVT leads to a significant depression of plasma LH perhaps due to an exhaustion of the releasable pool of LH in the anterior pituitary.
Assuntos
Ciproterona/farmacologia , Hormônio Liberador de Gonadotropina/farmacologia , Hormônio Luteinizante/sangue , Vasotocina/farmacologia , Animais , Arginina Vasopressina/farmacologia , Castração , Hormônio Liberador de Gonadotropina/administração & dosagem , Masculino , Ocitocina/farmacologia , Adeno-Hipófise/metabolismo , Ratos , Fatores de Tempo , Vasotocina/administração & dosagemRESUMO
The N-methyl-N-nitrosourea (NMU) model of hormone-responsive rat mammary carcinogenesis was used to address the hypothesis that melatonin (Mel), the principle hormone of the pineal gland, inhibits tumorigenesis by acting as an anti-promoting rather than an anti-initiating agent. Daily late-afternoon injections of Mel (500 micrograms/day), restricted to the initiation phase of NMU mammary tumorigenesis, were ineffective in altering tumor growth over a 20-week period. When Mel treatment was delayed for 4 weeks after NMU and then continued through the remainder of the promotion phase, only tumor number was significantly lower than in controls. However, when Mel injections encompassed the entire promotion phase, both tumor incidence and number were significantly lower than in the controls. Although elimination of the endogenous Mel signal via pinealectomy promoted tumor growth, the effect was not statistically significant. Serum levels of estradiol and tumor estrogen receptor content were unaltered by either Mel or pinealectomy. While Mel treatment failed to affect circulating prolactin levels, pinealectomy caused a two-fold increase in serum prolactin. The estradiol-stimulated recrudescence of tumors following ovariectomy was completely blocked by either 20, 100 or 500 micrograms Mel/day or tamoxifen (20 micrograms/day). Thus, Mel appears to be an anti-promoting hormone that may antagonize the tumor-promoting actions of estradiol in this model of mammary tumorigenesis.