RESUMEN
One part of chemical space that is endowed with interesting biological properties is the area of the chalcones. With this review, we provide a comprehensive overview of the numerous in vivo animal studies on the antineoplastic potential of both natural and synthetic members of this flavonoid subclass (covering: up to mid-2011). The thus far identified modes of action of these compounds are also discussed. We hope that this overview may stimulate deeper investigations into the biochemical mechanisms by which chalcones exert their antineoplastic action. As a result, in the foreseeable future, chalcones may prove suitable lead molecules or early drug candidates for the prevention or treatment of various neoplastic diseases.
Asunto(s)
Antineoplásicos/uso terapéutico , Chalconas/uso terapéutico , Animales , Antineoplásicos/síntesis química , Antineoplásicos/química , Chalconas/síntesis química , Chalconas/química , Modelos Animales de Enfermedad , Hesperidina/análogos & derivados , Hesperidina/química , Hesperidina/uso terapéutico , Humanos , Relación Estructura-ActividadRESUMEN
Because invasion is, either directly or via metastasis formation, the main cause of death in cancer patients, development of efficient anti-invasive agents is an important research challenge. We have established a screening program for potentially anti-invasive compounds. The assay is based on organotypic confronting cultures between human invasive cancer cells and a fragment of normal tissue in three dimensions. Anti-invasive agents appeared to be heterogeneous with regard to their chemical nature, but plant alkaloids, polyphenolics and some of their synthetic congeners were well represented. Even within this group, active compounds were quite diverse: (+)-catechin, tangeretin, xanthohumol and other prenylated chalcones, 3,7-dimethoxyflavone, a pyrazole derivative, an isoxazolylcoumarin and a prenylated desoxybenzoin. The data gathered in this system are now applied in two projects. Firstly, structure-activity relationships are explored with computer models using an artificial neural network approach, based on quantitative structural descriptors. The aim of this study is the prediction and design of optimally efficient anti-invasive compounds. Secondly, the metabolism of orally ingested plant polyphenolics by colonic bacteria is studied in a simulator of the human intestinal microbial ecosystem (SHIME) and in human intervention trials. This method should provide information on the final bioavailability of the active compounds in the human body, with regard to microbial metabolism, and the feasibility of designing pre- or probiotics that increase the generation of active principles for absorption in the gastro-intestinal tract. The final and global aim of all these studies is to predict, synthesize and apply in vivo molecules with an optimal anti-invasive, and hence an anti-metastatic activity against cancer.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Flavonoides/farmacología , Invasividad Neoplásica/prevención & control , Metástasis de la Neoplasia/prevención & control , Neoplasias/tratamiento farmacológico , Fenoles/farmacología , Animales , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/metabolismo , Proliferación Celular/efectos de los fármacos , Flavonoides/química , Flavonoides/metabolismo , Humanos , Estructura Molecular , Fenoles/química , Fenoles/metabolismo , Plantas/química , PolifenolesRESUMEN
UNLABELLED: Tibolone is used in postmenopausal women to alleviate menopausal symptoms and to prevent osteoporosis, but it does not stimulate the endometrium and the breast. Up to date, little data are available on the effect of tibolone on breast cancer initiation and progression. OBJECTIVE: In the present in vitro study, we investigated the effect of tibolone and its metabolites (3alpha-OH tibolone, 3beta-OH tibolone, the Delta4 isomer and the sulphated isoform) on invasion of human breast cancer cells. METHODS: The effect on invasion was evaluated in the chick heart invasion assay using MCF-7/6 cells and in the collagen type I invasion assay using T47-D cells. Furthermore, the compounds were tested in aggregation and migration assays. RESULTS: We observed that, at a concentration of 100 microM, tibolone and its 3beta-OH metabolite possess anti-invasive activities in the two different invasion assays. However, this was neither due to effects on cell-cell adhesion nor on motility. In an attempt to probe the mechanism underlying the anti-invasive effect, we found that pro-MMP-9 release was markedly reduced in the supernatant of MCF-7/6 breast cancer cells treated with tibolone, 3alpha-OH tibolone and the Delta4 isomer but, interestingly, not with the sulphated metabolite. CONCLUSION: We conclude that tibolone and its 3beta-OH metabolite have an anti-invasive effect on the tested breast cancer cell lines in vitro. This effect on invasion is not correlated with an effect on cell-cell adhesion or motility but coincides with a decreased release of pro-MMP-9 in the medium.
Asunto(s)
Adenocarcinoma/metabolismo , Neoplasias de la Mama/metabolismo , Proliferación Celular/efectos de los fármacos , Moduladores de los Receptores de Estrógeno/farmacología , Norpregnenos/farmacología , Adenocarcinoma/patología , Neoplasias de la Mama/patología , Línea Celular Tumoral/efectos de los fármacos , Moduladores de los Receptores de Estrógeno/uso terapéutico , Femenino , Humanos , Metaloproteinasa 9 de la Matriz/biosíntesis , Invasividad Neoplásica , Norpregnenos/uso terapéutico , Osteoporosis Posmenopáusica/prevención & controlRESUMEN
Fourteen novel C-prenylated and O-allylated 1,3-diarylpropenones (chalcones) were synthesized by Claisen-Schmidt condensation reaction of C-prenylated/O-allylated acetophenones with appropriate aldehydes; twelve of these model chalcones were screened in an assay based on the confrontation of invasive human MCF-7/6 mammary carcinoma cells with fragments of normal embryonic chick heart in vitro. Out of the twelve chalcones tested, three were found to exhibit potent anti-invasive activity. Some of these chalcones and their precursor acetophenones were also tested for inhibition of initiation of lipid peroxidation in rat liver microsomes; a prenylated acetophenone carrying two methoxy groups and two free phenolic hydroxy functions was found to be a potential antioxidant.
Asunto(s)
Antineoplásicos/síntesis química , Chalcona/farmacología , Invasividad Neoplásica/prevención & control , Acetofenonas/farmacología , Animales , Antineoplásicos/farmacología , Antioxidantes/síntesis química , Antioxidantes/farmacología , Neoplasias de la Mama/patología , Chalcona/síntesis química , Embrión de Pollo , Técnicas de Cocultivo , Técnicas Químicas Combinatorias , Evaluación Preclínica de Medicamentos , Femenino , Humanos , Peroxidación de Lípido/efectos de los fármacos , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Miocardio/citología , Ratas , Relación Estructura-Actividad , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Invasion of carcinoma cells is the result of a disequilibrium between invasion promoter and invasion suppressor gene products (1). The E-cadherin/catenin complex is the most potent invasion suppressor at the cell membrane of epithelioid cells (2).This complex consists of E-cadherin, a transmembrane glycoprotein of 120 kDa, which is linked to the actin cytoskeleton via the catenins (3). Downregulation of the complex is a common feature in invasive carcinoma cells, and has been recognized at several levels, ranging from genomic mutations to functional deficiencies of an apparently intact complex (4). Cell aggregation assays have been set up to test the functionality of the complex in epithelioid tumor cells. Functional integrity of the complex is a prerequisite for cell-cell adhesion between epithelial cells, and measuring cell aggregation in vitro has thus become another elegant tool to study differences between invasive and noninvasive cell types.
RESUMEN
Invasion occurs when invasion promoter molecules outbalance the function of invasion suppressors (1). Examples of invasion promoters are cell-matrix adhesion molecules, extracellular proteases, and cell motility factors. In normal tissues, positional stability of the cells is maintained through the counteraction of these invasion promoters by invasion suppressors such as enzyme inhibitors and cell-cell adhesion molecules. Within this context, the interaction of the cancer cells with their surrounding extracellular matrix (ECM) is a determining factor. To study this cell-matrix interaction in vitro, several natural ECM types have initially been applied. Bone (2), salt-extracted cartilage (3), and amnion membrane (4) are examples of devitalized substrata that have been launched in the past to discriminate between invasive and noninvasive cells. Lack of homogeneity of these substrata often made interpretation of invasion difficult, and hampered the reproducibility of those assays (5). To overcome these drawbacks, reconstituted and hence more homogeneous ECMs were developed, and proposed as substrata to test invasiveness. Matrigel (6) (as described in Chapter 7 by Hall and Brooks), and employed also in the assay described in Chapter 8 by Hendrix et al.), Humatrix (7) and collagen type I (8) are today frequently used ECMs in invasion assays. It should, however, be noted that, although these preparations may contain cytokines and growth factors, they are unable to react to the confrontation by cancer cells as a living host tissue does.
RESUMEN
Tumors are microecosystems in which a continuous cross-talk between cancer cells and host cells decides on the invasive behavior of the tumor cell population as a whole (1). Both compartments secrete activating and inhibitory factors that modulate activities such as cell-extracellular matrix (ECM) interaction, cell-cell adhesion, remodeling of the ECM, and cell motility. For this reason, confrontations of cancer cells with a living normal host tissue in organ culture have been introduced by several groups: Wolff and Schneider in France (2), Easty and Easty in the United Kingdom (3), and Schleich in Germany (4). Embryonic chick heart fragments in organ culture maintain many histological features of their tissue of origin: They are composed of myocytes, fibroblasts, and endothelial cells, and their ECM contains fibronectin, laminin, and several collagen types. Moreover, the fragments remain contractile, and this activity allows the monitoring of their functional integrity during organ culture.
RESUMEN
Tangeretin, a molecule present in citrus fruits and in certain 'natural' menopausal medications, is an effective tumour growth and invasion inhibitor in vitro of human MCF 7/6 breast cancer cells. However, when added to the drinking water of MCF 7/6 tumour-bearing mice it neutralises the beneficial tumour-suppressing effect of tamoxifen. Tangeretin reduces the number of natural killer cells. This may explain why the beneficial suppressive effect of tangeretin on MCF 7/6 cell proliferation in vitro is completely counteracted in vivo.
Asunto(s)
Antineoplásicos Hormonales/antagonistas & inhibidores , Antineoplásicos/farmacología , Flavonas , Flavonoides/farmacología , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Tamoxifeno/antagonistas & inhibidores , Animales , Antineoplásicos Hormonales/uso terapéutico , Femenino , Interacciones Alimento-Droga , Ratones , Tamoxifeno/uso terapéuticoRESUMEN
Cytokines and other paracrine or autocrine factors functionally modulate the invasion-suppressor and signal-transducing E-cadherin/catenin complex. We have used conditioned medium from human squamous carcinoma COLO 16 cells (CM COLO 16) as a source of such factors to modulate the E-cadherin/catenin complex in human breast carcinoma MCF-7 cells. CM COLO 16 induces scattering of MCF-7/AZ, but not of MCF-7/6 cells on tissue culture plastic substratum, and reduces aggregation of MCF-7/AZ cells in suspension. Insulin-like growth factor I counteracts this reduction of aggregation. Confocal laser scanning microscopy of immunocytochemical stainings shows loss of the honeycomb pattern of E-cadherin, alpha-catenin and beta-catenin, and internalization of those elements. Cell surface biotinylation shows a decrease in membrane-bound E-cadherin. Immunoprecipitation and cell fractionation show that the composition of the complex is maintained. Interleukin-1, interleukin-6, granulocyte-monocyte colony stimulating factor, stem cell factor, scatter factor/hepatocyte growth factor and transforming growth factor-beta, added separately to MCF-7/AZ cells, could not mimic the effects of CM COLO 16. Neither could we find evidence that the 80 kDa extracellular fragment of E-cadherin is implicated in scattering of MCF-7/AZ cells. This fragment is present in CM COLO 16, but it is also produced by the MCF-7/AZ cells themselves, even at higher levels. Our data point toward cytoplasmic internalization induced by paracrine factors as one of the downregulating mechanisms for the E-cadherin/catenin complex.
Asunto(s)
Neoplasias de la Mama , Cadherinas/metabolismo , Carcinoma de Células Escamosas , Medios de Cultivo Condicionados/farmacología , Proteínas del Citoesqueleto/metabolismo , Neoplasias Cutáneas , Transactivadores , Biotinilación , Western Blotting , Cadherinas/análisis , Agregación Celular/efectos de los fármacos , Agregación Celular/fisiología , Fraccionamiento Celular , Proteínas del Citoesqueleto/análisis , Desmoplaquinas , Factor de Crecimiento de Hepatocito/farmacología , Humanos , Factor I del Crecimiento Similar a la Insulina/farmacología , Interleucina-1/farmacología , Interleucina-4/farmacología , Interleucina-6/farmacología , Proteínas de la Membrana/metabolismo , Microscopía de Contraste de Fase , Pruebas de Precipitina , Receptor ErbB-2/metabolismo , Factor de Crecimiento Transformador beta/farmacología , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismo , alfa Catenina , beta CateninaRESUMEN
BACKGROUND: Tamoxifen and the citrus flavonoid tangeretin exhibit similar inhibitory effects on the growth and invasive properties of human mammary cancer cells in vitro; furthermore, the two agents have displayed additive effects in vitro. In this study, we examined whether tangeretin would enhance tamoxifen's therapeutic benefit in vivo. METHODS: Female nude mice (n = 80) were inoculated subcutaneously with human MCF-7/6 mammary adenocarcinoma cells. Groups of 20 mice were treated orally by adding the following substances to their drinking water: tamoxifen (3 x 10(-5) M), tangeretin (1 x 10(-4) M), tamoxifen plus tangeretin (3 x 10(-5) M plus 1 x 10(-4) M), or solvent. RESULTS AND CONCLUSIONS: Oral treatment of mice with tamoxifen resulted in a statistically significant inhibition of tumor growth compared with solvent treatment (two-sided P = .001). Treatment with tangeretin did not inhibit tumor growth, and addition of this compound to drinking water with tamoxifen completely neutralized tamoxifen's inhibitory effect. The median survival time of tumor-bearing mice treated with tamoxifen plus tangeretin was reduced in comparison with that of mice treated with tamoxifen alone (14 versus 56 weeks; two-sided P = .002). Tangeretin (1 x 10(-6) M or higher) inhibited the cytolytic effect of murine natural killer cells on MCF-7/6 cells in vitro, which may explain why tamoxifen-induced inhibition of tumor growth in mice is abolished when tangeretin is present in drinking water. IMPLICATIONS: We describe an in vivo model to study potential interference of dietary compounds, such as flavonoids, with tamoxifen, which could lead to reduced efficacy of adjuvant therapy. In our study, the tumor growth-inhibiting effect of oral tamoxifen was reversed upon addition of tangeretin to the diet. Our data argue against excessive consumption of tangeretin-added products and supplements by patients with mammary cancer during tamoxifen treatment.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Anticarcinógenos/uso terapéutico , Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Flavonas , Flavonoides/farmacología , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Extractos Vegetales/farmacología , Tamoxifeno/uso terapéutico , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/biosíntesis , Animales , Anticarcinógenos/sangre , Antineoplásicos/uso terapéutico , Sinergismo Farmacológico , Femenino , Flavonoides/uso terapéutico , Regulación Neoplásica de la Expresión Génica , Células Asesinas Naturales/efectos de los fármacos , Ratones , Ratones Desnudos , Extractos Vegetales/uso terapéutico , Tamoxifeno/sangre , Células Tumorales CultivadasRESUMEN
The acquisition of invasiveness is a crucial step in the malignant progression of cancer. In cancers of the colon and of other organs the E-cadherin/catenin complex, which is implicated in homotypic cell-cell adhesion as well as in signal transduction, serves as a powerful inhibitor of invasion. We show here that one allele of the alphaE-catenin (CTNNA1) gene is mutated in the human colon cancer cell family HCT-8, which is identical to HCT-15, DLD-1 and HRT-18. Genetic instability, due to mutations in the HMSH6 (also called GTBP) mismatch repair gene, results in the spontaneous occurrence of invasive variants, all carrying either a mutation or exon skipping in the second alphaE-catenin allele. The alphaE-catenin gene is therefore, an invasion-suppressor gene in accordance with the two-hit model of Knudsen for tumour-suppressor genes.
Asunto(s)
Neoplasias del Colon/genética , Proteínas del Citoesqueleto/genética , Proteínas de Unión al ADN/genética , Genes Supresores de Tumor/fisiología , Invasividad Neoplásica/genética , Alelos , Neoplasias del Colon/patología , Exones/genética , Humanos , Cariotipificación , Fenotipo , Mutación Puntual , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo , Células Tumorales Cultivadas , alfa CateninaRESUMEN
Invasiveness, the ability of certain tumour cells to migrate beyond their natural tissue boundaries, often leads to metastasis, and usually determines the fatal outcome of cancer. The need for anti-invasive agents has led us to search for possibly active compounds among alkaloids and polyphenolics. One hundred compounds were screened in an assay based on the confrontation of invasive human MCF-7/6 mammary carcinoma cells with fragments of normal embryonic chick heart in vitro. Anti-invasive activity was frequently found among chalcones having a prenyl group. Six compounds were found to inhibit invasion when added to the culture medium at concentrations as low as 1 microM. For at least three of them the anti-invasive effect could be associated with a cytotoxic effect on the MCF-7/6 cells, but not on the heart tissue. This selective cytotoxicity was substantiated by different methods, such as histology and growth assays (volume measurements, cell counts, MTT and sulforhodamine B assays). The anti-invasive effects of the compounds could neither be ascribed to induction of apoptosis nor to the promotion of cell-cell adhesion. Our data indicate that among the alkaloids and polyphenolics a number of molecules can inhibit growth and invasion of human mammary cancer cells via selective cytotoxicity.
Asunto(s)
Alcaloides/farmacología , Antineoplásicos Fitogénicos/farmacología , Invasividad Neoplásica , Fenoles/farmacología , Alcaloides/química , Animales , Antineoplásicos Fitogénicos/química , Embrión de Pollo , Flavonoides/química , Flavonoides/farmacología , Corazón/efectos de los fármacos , Humanos , Modelos Moleculares , Miocardio/citología , Fenoles/química , Relación Estructura-Actividad , Células Tumorales Cultivadas/efectos de los fármacosAsunto(s)
Cadherinas/genética , Neoplasias del Colon/patología , Proteínas del Citoesqueleto/genética , Mutación/genética , Transactivadores , Adhesión Celular/genética , Moléculas de Adhesión Celular/genética , Células Clonales , Codón/genética , Neoplasias del Colon/genética , Desmoplaquinas , Humanos , Invasividad Neoplásica , ARN Mensajero/genética , Células Tumorales Cultivadas , alfa Catenina , beta CateninaRESUMEN
When epithelial cells reach confluency in vitro, a number of energy-requiring activities such as growth and motility are contact-inhibited. We investigated the possible role of the E-cadherin/catenin complex, which acts as an invasion suppressor, in contact inhibition. Three strategies for modulation of the complex were used. Firstly, the cell-cell adhesion and signal transduction functions of E-cadherin were neutralized immunologically in human MCF-7/6 mammary carcinoma cells possessing a complete complex. Secondly, the effect of E-cadherin transfection in E-cadherin negative cell lines was investigated. Thirdly, alpha-catenin deficient variants of the human HCT-8/S11 colon carcinoma cell line were compared with their parent cells. In confluent cultures functional downregulation of the E-cadherin/catenin complex did not alter cell growth nor saturation density. This was shown by cell number counts, protein staining assays, cell cycle analysis, proliferation markers (Ki67 and Proliferating Cell Nuclear Antigen) and apoptosis assays. However, confluent cells with a functionally deficient complex showed positional instability and enhanced succinate dehydrogenase-mediated mitochondrial 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl) tetrazolium bromide (MTT) conversion, as compared to cells with an active complex. Our data indicate that contact inhibition of motility and of mitochondrial enzyme activity, but not of growth is regulated by the E-cadherin/catenin complex in epithelial cells.
Asunto(s)
Cadherinas/metabolismo , Proteínas del Citoesqueleto/metabolismo , Regulación hacia Abajo , Células Epiteliales/citología , Células Epiteliales/metabolismo , Cadherinas/genética , Recuento de Células , División Celular , Movimiento Celular , Humanos , Mitocondrias/metabolismo , Células Tumorales Cultivadas , alfa CateninaAsunto(s)
Cadherinas/fisiología , Proteínas del Citoesqueleto/fisiología , Flavonas , Invasividad Neoplásica , Metástasis de la Neoplasia , Transactivadores , Adenocarcinoma/patología , Animales , Neoplasias de la Mama/patología , Cadherinas/química , Polaridad Celular , Proteínas del Citoesqueleto/química , Citoesqueleto/metabolismo , Desmoplaquinas , Perros , Epitelio/metabolismo , Proteínas Fetales/metabolismo , Flavonoides/farmacología , Regulación Neoplásica de la Expresión Génica , Humanos , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/farmacología , Sustancias Macromoleculares , Familia de Multigenes , Invasividad Neoplásica/prevención & control , Metástasis de la Neoplasia/prevención & control , Proteínas de Neoplasias/fisiología , Conformación Proteica , Receptor IGF Tipo 1/efectos de los fármacos , Receptor IGF Tipo 1/fisiología , Relación Estructura-Actividad , Tamoxifeno/farmacología , Tretinoina/farmacología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/metabolismo , alfa Catenina , beta CateninaRESUMEN
All-trans-retinoic acid (RA), like insulin-like growth factor I (IGF-I) and tamoxifen, inhibit invasion of human MCF-7/6 mammary cancer cells in vitro. For tamoxifen and for IGF-I, activation of the invasion-suppressor function of the E-cadherin/catenin complex was shown to be the most probable mechanism of the anti-invasive action. We did a series of experiments to determine whether the anti-invasive effect of RA also implicated the invasion-suppressor E-cadherin/catenin complex. Human MCF-7/6 mammary and HCT-8/R1 colon cancer cells, both with a dysfunctional E-cadherin/catenin complex, were treated with RA and the function of the complex was evaluated through Ca(2+)-dependent fast aggregation. Fast aggregation of both MCF-7/6 and HCT-8/R1 cells was induced by 1 microM RA. This effect was abolished by antibodies against E-cadherin. RA-induced fast aggregation was not sensitive to cycloheximide, tyrosine kinase inhibitors or antibodies against IGF-I or against the IGF-I receptor. RA did not stimulate IGF-I receptor phosphorylation or alter the E-cadherin/catenin complex, as evidenced by immunoprecipitation. RA up-regulates the function of the invasion-suppressor complex E-cadherin/catenin. Its action mechanism is different from that of IGF-I. RA may act as an anti-invasive agent with unique mechanisms of action.
Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Cadherinas/fisiología , Tretinoina/farmacología , Cadherinas/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Agregación Celular/efectos de los fármacos , Humanos , Invasividad Neoplásica , Células Tumorales CultivadasRESUMEN
Loss of epithelioid organization in carcinoma cell lines has been related to invasiveness and poor differentiation of tumors. We investigated the invasion in vitro of various human colon cancer cell lines. Most cell lines were noninvasive into chick heart fragments, and this correlated with an epithelioid morphotype. Only cell lines COLO320DM, SW620, and variants of HCT-8 and DLD-1 were invasive and nonepithelioid. We examined in these cell lines whether invasiveness was related to changes in the structure and function of the E-cadherin/catenin complex. E-cadherin functions as an invasion suppressor and as a cell-cell adhesion molecule when linked to the cytoskeleton via alpha-catenin plus beta- or gamma-catenin. All noninvasive cell lines showed E-cadherin linked to these catenins. The E-cadherin-dependent cell-cell adhesion function in these cell lines was demonstrated by two assays in vitro. It was interesting that all invasive cell lines showed a dysfunctional E-cadherin/catenin complex. COLO320DM, SW480, and SW620 cells were defective in E-cadherin expression, whereas the invasive variants of HCT-8 and DLD-1 lacked the alpha-catenin protein. From clonal epithelioid HCT-8 cultures with functional E-cadherin/catenin complexes, we subcloned, repeatedly, round cell variants that were again invasive and expressed no alpha-catenin protein. Our data suggest that reproducible transformations toward a more invasive phenotype in HCT-8 cells are associated with down-regulation of alpha-catenin. The mechanisms of this transformation and the level of alpha-catenin down-regulation are currently investigated.
Asunto(s)
Neoplasias del Colon/patología , Proteínas del Citoesqueleto/metabolismo , Invasividad Neoplásica , Secuencia de Aminoácidos , Animales , Cadherinas/metabolismo , Calcio/metabolismo , Agregación Celular , Embrión de Pollo , Neoplasias del Colon/metabolismo , Epitelio/patología , Humanos , Técnicas Inmunológicas , Técnicas In Vitro , Datos de Secuencia Molecular , Péptidos/química , Péptidos/inmunología , Células Tumorales Cultivadas , alfa CateninaRESUMEN
MCF-7 human breast cancer cells express E-cadherin and show, at least in some circumstances, E-cadherin-dependent cell-cell adhesion (Bracke et al., 1993). The MCF-7/AZ variant spontaneously displays E-cadherin-dependent fast aggregation; in the MCF-7/6 variant, E-cadherin appeared not to be spontaneously functional in the conditions of the fast aggregation assay, but function could be induced by incubation of the suspended cells in the presence of insulinlike growth factor I (IGF-I) (Bracke et al., 1993). E-cadherin from MCF-7 cells was shown to contain sialic acid. Treatment with neuraminidase was shown to remove this sialic acid, as well as most of the sialic acid present at the cell surface. Applied to MCF-7/AZ, and MCF-7/6 cells, pretreatment with neuraminidase abolished spontaneous as well as IGF-I induced, E-cadherin-dependent fast cell-cell adhesion of cells in suspension, as measured in the fast aggregation assay. Treatment with neuraminidase did not, however, inhibit the possibly different, but equally E-cadherin-mediated, process of cell-cell adhesion of MCF-7 cells on a flat plastic substrate as assessed by determining the percentage of cells remaining isolated (without contact with other cells) 24 h after plating.
Asunto(s)
Cadherinas/metabolismo , Adhesión Celular , Ácidos Siálicos/metabolismo , Neoplasias de la Mama , Agregación Celular , Membrana Celular/metabolismo , Femenino , Humanos , Ácido N-Acetilneuramínico , Células Tumorales CultivadasRESUMEN
Tamoxifen is an antiestrogen used in adjuvant therapy of breast carcinoma and could potentially prevent the development of mammary cancer. While it is widely clinically used, its exact mechanisms of action are not yet fully elucidated. MCF-7/6 cells are estrogen receptor-positive invasive human breast cancer cells with a functionally inactive cell surface E-cadherin. In this study, we report that tamoxifen, and to a lesser extent its metabolites 4-OH-tamoxifen and N-desmethyltamoxifen, restore the function of E-cadherin in MCF-7/6 cells. In an aggregation assay, 10(-6) M tamoxifen significantly increases the aggregation of MCF-7/6 cells. This effect is abrogated by a monoclonal antibody against E-cadherin (HECD-1), is fast (within 30 min), and does not require de novo protein synthesis. Tamoxifen was also found to inhibit the invasion of MCF-7/6 cells in organ culture. Our data is the first demonstration that tamoxifen can activate the function of an invasion suppressor molecule and suggest that the restoration of E-cadherin function may contribute to the therapeutic benefit of tamoxifen in breast cancer patients.