RESUMEN
BACKGROUND: Adipose tissue transplantation has the benefit of providing both regenerative and aesthetic outcomes in breast cancer treatment. However, the transplanted tissue can stimulate the growth of residual cancer cells. OBJECTIVES: The aim of this study is to identify the interactions between adipose tissue cell subpopulations and human cancer cell lines. METHODS: Intact adipose tissue from lipofilling procedures as well as fibroblasts derived from adipose tissue, were cocultured in the presence of MDA-MB-231, MCF-7 e ZR-75-1 breast cancer cell lines. The influence on cancer cell lines of fibroblasts, induced to differentiate into specific adipocytes, was also assayed. RESULTS: All cancer cell lines displayed a significant increase in proliferation rate when cocultured in the presence of either intact adipose tissue or induced adipocytes. To a lesser extent, uninduced fibroblasts stimulate breast cancer cell proliferation. CONCLUSIONS: Recent studies have shown that the microenvironment surrounding breast cancer cells may stimulate growth and promote progression of residual cancer cells when surgery is performed on the main tumor mass. Accordingly, the graft of adipose tissue could potentially promote or accelerate the development of a subclinical tumor or support its locoregional recurrence. Our data suggest that adipocytes have a remarkable influence on the proliferation of cancer cell lines. The oncological safety of the lipofilling procedure outcome is still debated; thus, further studies and consistent follow-up examination are needed.
Asunto(s)
Adipocitos/fisiología , Adipogénesis , Tejido Adiposo/citología , Neoplasias de la Mama/patología , Comunicación Celular , Transdiferenciación Celular , Fibroblastos/fisiología , Adipocitos/metabolismo , Tejido Adiposo/trasplante , Adulto , Neoplasias de la Mama/metabolismo , Proliferación Celular , Técnicas de Cocultivo , Femenino , Fibroblastos/metabolismo , Humanos , Células MCF-7 , Persona de Mediana Edad , FenotipoRESUMEN
Sertoli cells play an essential role in the development of a functional testis. ROS (reactive oxygen species) are normally produced by the developing testicular cells and may be dangerous to spermatogenesis. The aim of this study was to investigate the developmental expression of genes involved in antioxidant defense as well as in the DNA damage response in rat Sertoli cells. As revealed by quantitative RT-PCR analysis, the expression pattern of the antioxidant enzymes GST (glutathione-S-transferase), CAT (catalase) and SOD (superoxide dismutase) showed a progressive decrease from birth to puberty. The expression level of the oncosuppressor p53 revealed a net reduction as well. We next focused on PARP-1 [poly(ADP-ribose) polymerase-1], a 'guardian of the genome' that combats stress conditions. At both the mRNA and protein level, PARP-1 expression was low at the early stage of development and increased later on. Maximal PARP-1 expression was preceded by a rise in the transcript level for MTs (metallothioneins), which provide zinc to zinc-dependent enzymes and proteins, including PARP-1. Our results showed an increased expression of PARP-1 during Sertoli cell development, together with a decrease in the expression of antioxidant enzymes. In conclusion, a role of PARP-1 in protecting the testicular differentiation is suggested.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Poli(ADP-Ribosa) Polimerasas/metabolismo , Células de Sertoli/metabolismo , Animales , Antioxidantes , Catalasa/genética , Catalasa/metabolismo , Diferenciación Celular , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Masculino , Poli(ADP-Ribosa) Polimerasa-1 , Poli(ADP-Ribosa) Polimerasas/genética , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Células de Sertoli/citología , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
The effect of growth hormone (GH) and cadmium (Cd) on metallothionein (MT) expression was investigated in hepatoma cells. In fish the constitutive isoform MT-B and the metal-responsive MT-A are expressed. Real-time RT-PCR revealed that: Cd up-regulates mostly MT-A, GH slightly induces MT-B and the GH/Cd combination induces synergistically both MTs. Perturbations in Ca2+ levels suppressed or reduced the Cd-induction of MTs and abolished the GH/Cd synergy. Similar results were obtained by inhibition of tyrosine kinases. Also the signaling molecules recruited by the GH receptor responded differently to GH and Cd, with ERKs showing a synergistic activation upon GH/Cd. The following conclusions can be drawn: (1) cytosolic Ca2+ is mainly involved in MT-A regulation; (2) both Ca2+ and tyrosine phosphorylation are essential for Cd-induction and GH/Cd synergy on MTs. The synergy could depend on interactions in different signaling pathways, leading to a differential recruitment of MTF-1 and AP-1 transcription factors.
Asunto(s)
Cadmio/farmacología , Carcinoma Hepatocelular/genética , Regulación de la Expresión Génica/efectos de los fármacos , Hormona del Crecimiento/farmacología , Metalotioneína/genética , Animales , Calcio/metabolismo , Carcinoma Hepatocelular/metabolismo , Proteínas de Unión al ADN/metabolismo , Sinergismo Farmacológico , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Metalotioneína/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Oncorhynchus mykiss/metabolismo , Fosforilación/efectos de los fármacos , Regiones Promotoras Genéticas , Unión Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Somatotropina/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción AP-1/metabolismo , Factores de Transcripción/metabolismo , Transcripción Genética , Células Tumorales Cultivadas , Tirosina/metabolismo , Factor de Transcripción MTF-1RESUMEN
The aim of the present study was to evaluate the in vivo effects of the RAR-alpha selective antagonist Ro 41-5253 on a xenograft animal model for breast cancer. Our observations indicate a lack of toxic side effects of the drug, even when used at high dosages. It is interesting to note that using Ro 41-5253 at dosages of 10, 30 and 100 mg/kg/die resulted in a slight, but significant inhibition of cell growth. The data obtained in this study represents the basis for a further evaluation of Ro 41-5253 anti-neoplastic activity on transgenic breast cancer animal models.
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Benzoatos/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Cromanos/uso terapéutico , Receptores de Ácido Retinoico/antagonistas & inhibidores , Retinoides/toxicidad , Animales , Línea Celular Tumoral , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Ratones , Receptor alfa de Ácido RetinoicoRESUMEN
PURPOSE: We assessed in vitro the corrosion behavior and biocompatibility of four Zr-based alloys (Zr97.5âNb1.5VM1.0ââ; VM, valve metal: Ti, Mo, W, Ta; at%) to be used as implant materials, comparing the results with grade-2 titanium, a biocompatible metal standard. METHODS: Corrosion resistance was investigated by open circuit potential and electrochemical impedance spectroscopy measurements as a function of exposure time to an artificial physiological environment (Ringer's solution). Human bone marrow stromal cells were used to evaluate biocompatibility of the alloys and their influence on growth kinetics and cell osteogenic differentiation through histochemical and gene expression analyses. RESULTS: Open circuit potential values indicated that Zr-based alloys and grade-2 Ti undergo spontaneous passivation in the simulated aggressive environment. High impedance values for all samples demonstrated improved corrosion resistance of the oxide film, with the best protection characteristics displayed by Zr97.5ââNb1.5Ta1.0. Cells seeded on all surfaces showed the same growth kinetics, although matrix mineralization and alkaline phosphatase activity were maximal on Zr97.5ââNb1.5Mo1.0 and Zr97.5âââNb1.5Ta1.0. Markers of ongoing proliferation, however, such as podocalyxin and CD49f, were still overexpressed on Zr97.5âââNb1.5âââMo1.0 even upon osteoinduction. No relevant effects were noted for the CD146-expressing population of bone progenitors. Nonetheless, the presence of a more differentiated cell population on Zr97.5Nb1.5Ta1.0 samples was inferable by comparing mineralization data and transcript levels of osteogenic markers (osteocalcin, osteopontin, bone sialoprotein, and RUNX2). CONCLUSIONS: The combination of passivation, corrosion resistance and satisfactory biotolerance to bone progenitors make the Zr-based alloys promising implant materials. Among those we tested, Zr97.5Nb1.5Ta1.0 seems to be the most appealing.
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Aleaciones , Prótesis Dental , Aparatos Ortopédicos , Titanio , Circonio , Aleaciones/química , Aleaciones/farmacología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Corrosión , Espectroscopía Dieléctrica/métodos , Humanos , Ensayo de Materiales/métodos , Células Madre Mesenquimatosas/efectos de los fármacos , Titanio/química , Titanio/farmacología , Circonio/química , Circonio/farmacologíaRESUMEN
Hormonal changes in humans during spaceflight have been demonstrated but the underlying mechanisms are still unknown. To clarify this point thyroid and testis/epididymis, both regulated by anterior pituitary gland, have been analyzed on long-term space-exposed male C57BL/10 mice, either wild type or pleiotrophin transgenic, overexpressing osteoblast stimulating factor-1. Glands were submitted to morphological and functional analysis.In thyroids, volumetric ratios between thyrocytes and colloid were measured. cAMP production in 10(-7)M and 10(-8)M thyrotropin-treated samples was studied. Thyrotropin receptor and caveolin-1 were quantitized by immunoblotting and localized by immunofluorescence. In space-exposed animals, both basal and thyrotropin-stimulated cAMP production were always higher. Also, the structure of thyroid follicles appeared more organized, while thyrotropin receptor and caveolin-1 were overexpressed. Unlike the control samples, in the space samples thyrotropin receptor and caveolin-1 were both observed at the intracellular junctions, suggesting their interaction in specific cell membrane microdomains.In testes, immunofluorescent reaction for 3ß- steroid dehydrogenase was performed and the relative expressions of hormone receptors and interleukin-1ß were quantified by RT-PCR. Epididymal sperm number was counted. In space-exposed animals, the presence of 3ß and 17ß steroid dehydrogenase was reduced. Also, the expression of androgen and follicle stimulating hormone receptors increased while lutenizing hormone receptor levels were not affected. The interleukin 1 ß expression was upregulated. The tubular architecture was altered and the sperm cell number was significantly reduced in spaceflight mouse epididymis (approx. -90% vs. laboratory and ground controls), indicating that the space environment may lead to degenerative changes in seminiferous tubules.Space-induced changes of structure and function of thyroid and testis/epididymis could be responsible for variations of hormone levels in human during space missions. More research, hopefully a reflight of MDS, would be needed to establish whether the space environment acts directly on the peripheral glands or induces changes in the hypotalamus-pituitary-glandular axis.
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Vuelo Espacial , Testículo/citología , Testículo/metabolismo , Glándula Tiroides/citología , Glándula Tiroides/metabolismo , Animales , Western Blotting , Caveolina 1/metabolismo , AMP Cíclico/biosíntesis , Técnica del Anticuerpo Fluorescente , Inmunohistoquímica , Interleucina-1beta/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Receptores de Esteroides/metabolismo , Receptores de Tirotropina/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Recuento de EspermatozoidesRESUMEN
Interference between heavy metals and growth hormone (GH) on cell signaling has been previously demonstrated in fish cells. This study was aimed at assessing their effects on expression of the metallothionein isoforms MT-A and MT-B. The results indicate that all heavy metals induce MT-A more markedly than MT-B, but differences appeared when metals were combined with GH. For MT-B induction, a positive interference between metals and GH was observed for Zn(2+)/GH and Cd(2+)/GH, a negative interference for Hg(2+)/GH. With regards to MT-A, no interference was observed for Zn(2+)/GH and Hg(2+)/GH, while a negative interference occurred with Cu(2+)/GH and a positive interference with Cd(2+)/GH. The possible mechanisms underlying the differential regulation of metallothioneins include different signaling pathways. The results show that STAT5 and ERKs responded differently to different combinations, and Zn(2+)/GH and Cd(2+)/GH exerted a slight positive interference on ERK activation. On the other hand, a synergic rise in [Ca(2+)](i) occurred for all combinations except for Cu(2+)/GH. Our data suggest that the cross-talk between heavy metals and GH resulting in MT transcription modulation does not strictly depend on Ca(2+) signalling; (ii)ERK activation may represent the point of cross-talk between Zn(2+) or Cd(2+) and GH, converging on MT-B transcription, probably through a differential recruitment of transcription factors.
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Metalotioneína/metabolismo , Metales Pesados/metabolismo , Animales , Cadmio/metabolismo , Calcio/metabolismo , Línea Celular , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Hormona del Crecimiento/metabolismo , Hormona del Crecimiento/farmacología , Metalotioneína/biosíntesis , Metales Pesados/farmacología , Oncorhynchus mykiss , Fosforilación , Factor de Transcripción STAT5/metabolismo , Zinc/farmacologíaRESUMEN
There is strong evidence that thyroid hormones through triiodothyronine (T3) regulate Sertoli cell proliferation and differentiation in the neonatal testis. However, the mechanism(s) by which they are able to control Sertoli cell proliferation is unclear. In the present study in vivo approaches (PTU-induced neonatal hypothyroidism known to affect Sertoli cell proliferation) associated with in vitro experiments on a Sertoli cell line were developed to investigate this question. We demonstrated that the inhibitory effect of T3 on Sertoli cell growth, analyzed by evaluating DNA-incorporated [3H] thymidine, was associated with a time and dose-dependent increase in the levels of Cx43, a constitutive protein of gap junctions, known to participate in the control of cell proliferation and the most predominant Cx in the testis. These Cx43 changes were associated with increased gap junction communication measured by gap FRAP. Consistent with these results two specific inhibitors of gap junction coupling, AGA and oleamide, were able to significantly reverse the T3 inhibitory effect on Sertoli cell proliferation. The present data also revealed a nongenomic effect of T3 on Cx43 Sertoli cells that was evidenced by a rapid up-regulation of gap junction plaque number as identified in Cx43-GFP transfected cells exposed to the hormone. This process appears mediated through actin cytoskeleton since incubation of the cells with cytochalasin D totally reversed the T3 stimulatory effect on Cx43-GFP gap junction plaques. Based on these data, we propose a working hypothesis in which Cx43 could be an intermediate target for T3 inhibition of neonatal Sertoli cell proliferation.
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Conexina 43/metabolismo , Uniones Comunicantes/fisiología , Células de Sertoli/fisiología , Triyodotironina/farmacología , Animales , Animales Recién Nacidos , Línea Celular , Proliferación Celular , Masculino , Ratones , Ratones Transgénicos , Feniltiourea/análogos & derivados , Ratas , Ratas Wistar , Túbulos Seminíferos/metabolismo , Hormonas Tiroideas/farmacología , TransfecciónRESUMEN
Malignant mesothelioma (MM) still remains a therapeutic and diagnostic problem to which new therapeutic perspectives are being continuously tried and tested. Three different primary cultures (MMGe-1, MES MM 98, and MES 1) and one immortalized cell line (MSTO 211 H) of human MM were studied in order to evaluate the HER-2/neu expression. Three out of four cell lines showed a different level of c-erbB-2 expression, the highest being detected on the MSTO 211 H cell line (fibroblastic phenotype), whereas MMGe-1 resulted negative. The effect of the anti-HER-2/neu antibody (Trastuzumab) alone, and in combination with cisplatin (CDDP) at different doses (ranging from 0.1 to 100 microg/ml), was studied on all the c-erB-2 positive cell lines. Trastuzumab was able to inhibit cell proliferation in a time-dependent manner, with growth inhibition also obtained at low concentrations (0.1-1 microg/ml). Combined treatment with Trastuzumab (10 microg/ml) and CDDP (1 microg/ml) showed synergism. Our results were encouraging, and suggest a rationale for further investigations in a clinical setting.