Expresión de receptores a factor de crecimiento insulínico tipo I, c-fos y PCNA en un tumor mamario experimental / Expression to an insulin growth factor type I receptor, c-fos and Nuclear Antigen of cellular proliferatium in an experimental mammary tumor

Las terapias oncológicas conllevan generalmente efectos secundarios indeseados por lo que el mejor conocimiento de los mecanismos regulatorios del desarrollo y crecimiento tumoral puede abrir el camino a enfoques terapeúticos más adecuados. El objetivo de éste trabajo fue profundizar el estudio de la implicancia de factores que regulan el crecimiento del cáncer mamario empleando un modelo experimental químicamente inducido en rata, el que presenta similitudes con el cáncer mamario humano principalmente en lo que respecta a la regulación hormonal de su crecimiento. El tumor mamario fue inducido químicamente en ratas normales y diabéticas. Se analizó la expresión de receptores a factor de crecimiento insulínico tipo I (RIGF-I), el que forma parte de un sistema formado por factores de crecimiento, sus receptores y proteínas transportadas; éste sistema se encuentra alterado en pacientes con diabetes mellitus no dependiente de insulina. También se analizó la expresión de las proteínas c-FOS y PCNA (antígeno nuclear de proliferación celular), ambas relacionadas con la proliferación celular. Los resultados experimentales mostraron significativas diferencias en los tumores mamarios desarrollados: los de las ratas diabéticas presentaron mayor período de latencia (p<0,001), menor número de tumores desarrollados por rata (p<0,02) y una velocidad de crecimiento menor (p<0,05) con respecto a los tumores desarrollados en ratas normales. Asimismo, mostraron un patrón histológico de marcada benignidad, en contraste con los adenocarcinomas malignos ductales desarrollados en los animales normales. La expresión de las proteínas c-FOS y PCNA detectada por métodos inmunohistoquímicos fue significativamente menor en los tumores de las ratas diabéticas que en ratas normales. En cuanto a la expresión de RIGF-I, los resultados indicaron que la misma estaría regulada por las hormonas esteroides en animales diabéticos y normales. El trabajo permitió analizar experimentalmente la interrelación entre factores de crecimiento insulínicos y hormonas esteroides en el desarrollo y crecimiento tumoral mamario, particularmente cuando están presentes la patología mamaria y la diabetes

Expresión de receptores a factor de crecimiento insulínico tipo I, c-fos y PCNA en un tumor mamario experimental / Expression to an insulin growth factor type I receptor, c-fos and Nuclear Antigen of cellular proliferatium in an experimental mammary tumor

Las terapias oncológicas conllevan generalmente efectos secundarios indeseados por lo que el mejor conocimiento de los mecanismos regulatorios del desarrollo y crecimiento tumoral puede abrir el camino a enfoques terapeúticos más adecuados. El objetivo de éste trabajo fue profundizar el estudio de la implicancia de factores que regulan el crecimiento del cáncer mamario empleando un modelo experimental químicamente inducido en rata, el que presenta similitudes con el cáncer mamario humano principalmente en lo que respecta a la regulación hormonal de su crecimiento. El tumor mamario fue inducido químicamente en ratas normales y diabéticas. Se analizó la expresión de receptores a factor de crecimiento insulínico tipo I (RIGF-I), el que forma parte de un sistema formado por factores de crecimiento, sus receptores y proteínas transportadas; éste sistema se encuentra alterado en pacientes con diabetes mellitus no dependiente de insulina. También se analizó la expresión de las proteínas c-FOS y PCNA (antígeno nuclear de proliferación celular), ambas relacionadas con la proliferación celular. Los resultados experimentales mostraron significativas diferencias en los tumores mamarios desarrollados: los de las ratas diabéticas presentaron mayor período de latencia (p<0,001), menor número de tumores desarrollados por rata (p<0,02) y una velocidad de crecimiento menor (p<0,05) con respecto a los tumores desarrollados en ratas normales. Asimismo, mostraron un patrón histológico de marcada benignidad, en contraste con los adenocarcinomas malignos ductales desarrollados en los animales normales. La expresión de las proteínas c-FOS y PCNA detectada por métodos inmunohistoquímicos fue significativamente menor en los tumores de las ratas diabéticas que en ratas normales. En cuanto a la expresión de RIGF-I, los resultados indicaron que la misma estaría regulada por las hormonas esteroides en animales diabéticos y normales. El trabajo permitió analizar experimentalmente la interrelación entre factores de crecimiento insulínicos y hormonas esteroides en el desarrollo y crecimiento tumoral mamario, particularmente cuando están presentes la patología mamaria y la diabetes (AU)

Histamine as an autocrine growth factor: an unusual role for a widespread mediator.

The involvement of histamine in cancer growth represents an old controversy and direct experimental evidence proving this hypothesis is not still available. In this paper we review the most relevant mechanisms referring to the role of histamine receptors, histidine decarboxylase and histamine release in the onset of an autocrine loop, that enables histamine to act as an autocrine growth factor. We postulate that this autocrine loop, that has been studied in an experimental mammary carcinoma model induced in rats, may be present in different human neoplasias. Therefore, the better understanding of this novel regulatory pathway that is controlled by histamine may contribute to identifying new therapeutic targets.

An experimental model of diabetes and cancer in rats.

The aim of this study was to develop an experimental model for the study of cancer associated with diabetes. For diabetes induction, Sprague-Dawley rats were given streptozotocin (STZ, 90 mg/kg body weight (BW), by intraperitoneal injection on the second day of life. For mammary tumour induction, rats were injected with 50 mg/kg BW of N-nitroso-N-methylurea (NMU) at 50, 80 and 110 days old. The neoplastic process and the effect of tamoxifen treatment was examined in non-diabetic and diabetic rats. The latency period, NMU-induced tumour incidence and the number of tumours per rat in diabetic rats versus controls were 117 +/- 7 days versus 79 +/- 9 days (P < 0.001); 93% versus 95% (NS); and 5.2 +/- 1.6 versus 2.7 +/- 0.5 (P < 0.02). A more benign histological pattern for tumours in diabetic animals was observed. Mammary tumours in diabetic rats grew more slowly than in controls. Tamoxifen (1 mg/kg/day) treated diabetic rats showed tumour regression in 67% of NMU-induced mammary tumours versus 53% in controls (NS). Our results show that tumour progression seems to be affected by diabetes in this experimental model. We suggest this is the result of changes to insulin-like growth factors and their receptors, which occur in diabetics, and our future research will examine this hypothesis.

Epidermal growth factor in NMU-induced mammary tumors in rats.

In this work we analyze the hypothesis that tumors induced by i.p. N-nitroso-N-methylurea injection express EGF-like peptides and EGF receptors which could be involved in the response to hormone manipulation. EGF receptors (EGFR) were determined in the purified membrane fraction of tumors from control and ovariectomized (OVX) animals and no significant differences were found in either maximal binding capacities (Q) or dissociation constants (Kd) between them. Neither did we observe differences between tumors that regressed (HR) or continued growing (HU) after ovariectomy. In order to test the ability of EGFR to trigger a biological response we measured the production of second messengers inositol triphosphates (IP3) and cAMP levels; we found that EGF increases IP3 production in a dose-dependent way, while cAMP levels were not affected. In addition, EGF was able to induce in vitro cell proliferation in a concentration-dependent manner when tested in primary cultures of tumor cells by the clonogenic soft agar technique. EGF/TGF-alpha activity was determined by a radioreceptor assay in tumor cytosols from control and OVX rats. Results showed a trend to lower values in tumors from OVX rats, but no differences between HR and HU tumors. A positive correlation was found between EGF/TGF-alpha activity and progesterone receptor maximal binding capacity. When we tested the action of estradiol and EGF added together to primary cultures of tumor cells we found an additive effect on cell proliferation. The study of steady state mRNA levels showed that E2 increases PgR and c-myc mRNA levels in HR but not in HU tumors. In conclusion, the autocrine loop EGFR-EGF/TGF-alpha present in all tumors is hormonally regulated, possibly by Pg, but is not related to the tumor response to ovariectomy.

Atypical association of H1 and H2 histamine receptors with signal transduction pathways during multistage mouse skin carcinogenesis.

OBJECTIVE: In the present work we studied the association of histamine receptors with second messengers during multistage carcinogenesis in Sencar mice skin. METHODS: 96 Sencar female mouse, divided into six groups were used. Tumors appeared only in the 7, 12-dimethylbenz[a]anthracene-initiated and 12-O-tetradecanoylphorbol-13-acetate-promoted group. Control groups received only TPA, or acetone or no treatment at all. Periodically during the promotion period, cAMP and inositol phosphate production were measured after stimulation with H1 or H2 agonists in samples from all groups. RESULTS: In non-treated skin, H1 receptors were coupled to phosphatidylinositol hydrolysis and H2 receptors mediated cAMP production. Conversely, in tumors H2 receptors were associated with phosphatidylinositol hydrolysis and H1 mediated a rise in cAMP levels. The skin among tumors and the skin from all control groups maintained the same coupling as non-treated skin. An increase in mast cell number, with a homogeneous subepithelial distribution and marked phenotypic changes, was also observed in promoted skin. CONCLUSIONS: These findings indicate an atypical association of histamine receptors with second messengers that could be a critical feature for the postulated action of histamine in tumor growth.

Receptors characterization of intraperitoneally N-nitroso-N-methylurea-induced mammary tumors in rats.

Mammary adenocarcinomas induces in female Sprague-Dawley rats by three intraperitoneal injections of N-nitroso-N-methylurea were studied in order to characterize their estrogen (ER), progesterone (PgR), prolactin (PRLR) and epidermal growth factor (EGFR) receptors. All samples evaluated showed the presence of ER and PgR in the cytosol fraction and PRLR amd EGFR in the membrane fraction. Q (fmol/mg) and K(d) (nM) values were as follows: ER, 56 +/- 11 and 0.5 +/- 0.1; PgR, 109 +/- 25 and 2.2 +/- 0.5 and PRLR, 335 +/- 75 and 0.5 +/- 0.2, respectively. In all tumors studied, two specific sites were found for EGFR, one with Q(1) = 22 +/- 9 and K(d1) = 0.6 +/- 0.3, and the other with Q(2) = 125 +/- 33 and K(d2) = 2.1 +/- 0.5. Receptor content was found to be independent of tumor histopathological variety. Displacement index (DI) with estradiol and tamoxifen of [I(3)H]E2-ER binding showed great heterogeneity, with values ranging from 0.01 to1.54. No correlation between ER content and DI values was found. Antiestrogenic binding sites were not found in the microsomal fraction of ten mammary tumors examined. Proliferation of this experimental mammary tumor may be regulated by a complex interaction of steroid and polypeptide hormones, as well as growth factors.

Effect of tamoxifen on intraperitoneal N-nitroso-N-methylurea induced tumors.

The effect of tamoxifen (TAM) was evaluated on a mammary tumor model induced in Sprague-Dawley rats by intraperitoneal administration of three N-nitroso-N-methylurea (NMU) doses. Animals received TAM (1 mg/kg per day) from 10 days before the first NMU dose up to 140 days later. Thereafter, treatment was discontinued and the observation period was extended 60 days longer. Mean overall latency period, tumor number per rat and tumor incidence were recorded. Significant differences between treated and control batches were observed in tumor number per rat (1.8 +/- 1.1 versus 5.2 +/- 1.6; P < 0.05) and in tumor incidence (50% versus 100%; P < 0.05), respectively. No significant difference in latency period between both batches was recorded. All lesions induced in the control batch were malignant, whereas only 45% of those induced in TAM-treated animals were malignant and the remaining 55% were preneoplastic. At 60 days after treatment discontinuance, tumor incidence increased to 90% and also tumor number per rat increased to 4.6 +/- 1.5. TAM effect was also evaluated in rats with NMU-induced tumors by treatment with 1 mg/kg per day during 60 days starting when tumors reached a 1.5-cm diameter. Regression to less than 80% of initial size in 49% of the tumors was observed, while in ovariectomized rats, 33% of tumors regressed. Estrogen receptor content, ER (fmol/mg protein) and Kd (nM) in control tumors were: 56 +/- 10 and 0.5 +/- 0.1. In tumors of TAM-treated animals, ER was less than 5 fmol/mg protein. Findings demonstrate that TAM significantly decreased the appearance of tumors induced in rats by i.p. injection of NMU and when TAM treatment was initiated after tumor induction, some tumors failed to respond to hormonal manipulation. Differential tumor growth response after TAM or oophorectomy in each tumor indicates that in the same rat it is possible to distinguish hormone-dependent and hormone-autonomous tumor populations. Hormonal regulation of tumor growth can be under intrinsic control, regardless of the hormonal status of the whole organism.