Long term inhibition by estradiol or progesterone of melatonin secretion after administration of a mammary carcinogen, the dimethyl benz(a)anthracene, in Sprague-Dawley female rat; inhibitory effect of Melatonin on mammary carcinogenesis.

Abstract

A single intragastric administration of 7,12-dimethylbenz(a)anthracene (DMBA) has been shown to induce mammary tumors in young cycling female Sprague-Dawley rats. The appearance of the tumors is preceded by a series of neuroendocrine disturbances, including attenuation of the preovulatory Luteinizing Hormone surge and Gonadotropin-Releasing Hormone release and amplification of the preovulatory 17beta-Estradiol (E2) surge. In this study, we examined the hypothesis that a single administration of DMBA increases the E2 and Progesterone inhibition of the spontaneous and Isoproterenol-induced Melatonin (MT) secretion from the pineal gland, during the latency phase. Also, the incidence of mammary tumors, as well as the possible preventive effect of various doses of Melatonin, were recorded up to 6 months after daily administration. For all studies, Sprague-Dawley rats, 55-60 days of age, received, on the Estrous day of the Estrous cycle, a single dose of 15 mg DMBA delivered by intragastric intubation. For the study on ovarian steroids, they were ovariectomized 5 days later and then sacrificed by decapitation at 10 a.m., one month later. Pineal glands were removed and placed in perifusion chambers containing Hanks 199 medium. The medium was saturated with O2/CO2 (95%/5%) and its pH was 7.4. Ten independent chambers were immersed in a water bath at 37 degrees C. Each pineal gland received medium (flow rate: 0.16 ml/min) through a system of input lines. The fractions were collected every 10 min, and immediately frozen at -20 degrees C until Melatonin RIA. Experiments were repeated to obtain up to five experimental points for each treatment. E2 (10(-11)-10(-9) M) and Progesterone (10(-9)-10(-7) M) were applied during the entire perifusion period (7 h). Isoproterenol (10(-6) M) was applied for 20 min after 2.5 h in perifusion. Melatonin concentrations and Areas Under the Curves were compared using two-factor ANOVA as well as parametric or nonparametric two-sample methods after testing sample normality. For the study on the possible preventive effect of Melatonin, they were daily treated, by the intragastric route, with increasing doses of Melatonin for 6 months. The percentage of female rats having at least one mammary carcinoma were compared using the Fischer exact t-test. During the latency phase, in vehicle-treated rats, E2 and Progesterone treatments lead an almost significant inhibition of the Isoproterenol-induced stimulation of Melatonin secretion. In DMBA-treated rats, E2 treatment leads to a complete blunting of the Isoproterenol-induced stimulation of Melatonin and Progesterone treatment leads to a cyclic inhibition of the Isoproterenol-induced Melatonin secretion. During the promotion phase, there was a dose-dependent inhibitory effect (up to 65% inhibition) of the daily administration of Melatonin, on mammary tumors occurrence. In conclusion, the long term inhibition of DMBA upon Melatonin secretion from the pineal gland might accelerate the promotion of mammary tumors induced by the mammary carcinogen. Inversely, the daily administration of Melatonin for 6 months induces a long lasting protective effect against the formation of mammary tumors.