RESUMEN
Endocrine disrupting chemicals (EDCs) are associated with cancer development and progression due to their promotion of increased cell invasiveness and metastasis formation. However, the effects of EDCs on cell adhesion mediated through integrins have not been well studied to date. Their actions are implicated by binding sites for hormones on the vitronectin receptor (VTNR; or integrin αvß3), which is involved in tumor angiogenesis and metastasis. VTNR-expressing human umbilical vein endothelial cells (HUVECs) were used to determine the effects of EDCs and endogenous hormones on cell adhesion to vitronectin-coated surfaces, and on VTNR activation. Cell adhesion was significantly increased for bisphenol A, triclocarban, and triclosan (10, 100 nM; p < 0.05), with similar trends for bisphenols AF and S (10, 100 nM; p > 0.05). No changes in cell adhesion were seen for 5α-dihydrotestosterone, 17ß-estradiol, triiodothyronine, imatinib and paroxetine. These data indicate that EDC-mediated increases in HUVEC adhesion to vitronectin are not mediated through androgenic, estrogenic, or thyroid activities, nor through activation of VTNR. Although these effects of EDCs on HUVEC adhesion require further investigation of the underlying mechanism(s) of action to define their biological relevance, the low-dose effects and nonmonotonic responses revealed here define the need for further investigation of these EDCs.
Asunto(s)
Adhesión Celular/efectos de los fármacos , Disruptores Endocrinos/toxicidad , Integrina alfaVbeta3/efectos de los fármacos , Compuestos de Bencidrilo/toxicidad , Carbanilidas/toxicidad , Fluorocarburos/toxicidad , Hormonas/toxicidad , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Humanos , Fenoles/toxicidad , Sulfonas/toxicidad , Triclosán/toxicidad , Vitronectina/metabolismoRESUMEN
RATIONALE: A high risk of post-operative recurrence contributes to the poor prognosis and low survival rate of oesophageal squamous cell carcinoma (ESCC) patients. Increasing experimental evidence suggests that integrin adhesion receptors, in particular integrin αv (ITGAV), are important for cancer cell survival, proliferation and migration. Therefore, targeting ITGAV may be a rational approach for preventing ESCC recurrence. MATERIALS AND METHODS: Protein levels of ITGAV were determined in human ESCC tumour tissues using immunohistochemistry. MTT, propidium iodide staining, and annexin V staining were utilized to investigate cell viability, cell cycle progression, and induction of apoptosis, respectively. Computational docking was performed with the Schrödinger Suite software to visualize the interaction between indomethacin and ITGAV. Cell-derived xenograft mouse models, patient-derived xenograft (PDX) mouse models, and a humanized mouse model were employed for in vivo studies. RESULTS: ITGAV was upregulated in human ESCC tumour tissues and increased ITGAV protein levels were associated with poor prognosis. ITGAV silencing or knockout suppressed ESCC cell growth and metastatic potential. Interestingly, we identified that indomethacin can bind to ITGAV and enhance synovial apoptosis inhibitor 1 (SYVN1)-mediated degradation of ITGAV. Integrin ß3, one of the ß subunits of ITGAV, was also decreased at the protein level in the indomethacin treatment group. Importantly, indomethacin treatment suppressed ESCC tumour growth and prevented recurrence in a PDX mouse model. Moreover, indomethacin inhibited the activation of cytokine TGFß, reduced SMAD2/3 phosphorylation, and increased anti-tumour immune responses in a humanized mouse model. CONCLUSION: ITGAV is a promising therapeutic target for ESCC. Indomethacin can attenuate ESCC growth through binding to ITGAV, promoting SYVN1-mediated ubiquitination of ITGAV, and potentiating cytotoxic CD8+ T cell responses.
Asunto(s)
Neoplasias Esofágicas/tratamiento farmacológico , Carcinoma de Células Escamosas de Esófago/tratamiento farmacológico , Indometacina/farmacología , Integrina alfaVbeta3/efectos de los fármacos , Integrina alfaVbeta3/metabolismo , Recurrencia Local de Neoplasia/prevención & control , Animales , Antiinflamatorios no Esteroideos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Xenoinjertos , Humanos , Ratones DesnudosRESUMEN
We previously demonstrated that F4 peptide (CNPEDCLYPVSHAHQR) from collagen XIX was able to inhibit melanoma cell migrationin vitro and cancer progression in a mouse melanoma model. The aim of the present work was to study the anti-angiogenic properties of F4 peptide. We demonstrated that F4 peptide inhibited VEGF-induced pseudo-tube formation on Matrigel by endothelial cells and endothelial sprouting in a rat aortic ring assay. By affinity chromatography, we identified αvß3 and α5ß1 integrins as potential receptors for F4 peptide on endothelial cell surface. Using solid phase assays, we proved the direct interaction between F4 and both integrins. Taken together, our results demonstrate that F4 peptide is a potent antitumor agent inhibiting both angiogenesis and tumor cell migration.
Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Colágeno/metabolismo , Integrina alfa5beta1/metabolismo , Integrina alfaVbeta3/metabolismo , Neovascularización Patológica/tratamiento farmacológico , Fragmentos de Péptidos/metabolismo , Animales , Antineoplásicos/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colágeno/farmacología , Células Endoteliales/metabolismo , Humanos , Integrina alfa5beta1/efectos de los fármacos , Integrina alfaVbeta3/efectos de los fármacos , Neovascularización Patológica/patología , Fragmentos de Péptidos/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To examine the effects of cisplatin on uterine histology and implantation molecules and the possible protective role of recombinant Klotho administration on uterine histology and uterine receptivity in mice exposed to cisplatin. MATERIALS AND METHODS: This study was conducted using thirty-two adult female mice assigned to four groups with 8 mice in each group. Saline was given to the 1st group, cisplatin to the 2nd group, recombinant mouse Klotho to the 3rd group and recombinant mouse Klotho plus cisplatin to the 4th group. Uterine tissues were examined for damage histologically and immunobiologically for the uterine receptivity markers HOXA13 and alphaVBeta3 integrin. RESULTS: Apoptosis, degeneration, decrease in uterine thickness and uterine absence of gland scores were higher in the cisplatin group (3rd group) compared to the saline group (1st group) (cisplatin vs. saline p < 0.0001 for all parameters). In the recombinant Klotho plus cisplatin group (4th group), scores of apoptosis, degeneration, reduction in uterine thickness and uterine absence of gland were lower than the group receiving only cisplatin (cisplatin plus recombinant Klotho vs cisplatin, p = 0.006 for apoptosis; p = 0.017 for degeneration; p = 0.011 for the reduction in uterine thickness; p = 0.002 for the absence of gland). However, HOXA13 and alphaVBeta3 integrin staining levels were not different between the cisplatin group (group 3) and the cisplatin plus recombinant Klotho group (group 4) (p = 0.980 and p = 0.762, respectively.) CONCLUSION: Cisplatin has adverse effects on the uterus. Administration of recombinant Klotho was found to attenuate the cisplatin-induced damage but failed to preserve levels of the implantation molecules HOXA13 and alphaVbeta3. Further studies examining the effect of cisplatin toxicity using other implantation markers along with functional studies are needed.
Asunto(s)
Antineoplásicos/efectos adversos , Cisplatino/efectos adversos , Implantación del Embrión/efectos de los fármacos , Proteínas de Homeodominio/efectos de los fármacos , Integrina alfaVbeta3/efectos de los fármacos , Útero/metabolismo , Animales , Femenino , Glucuronidasa/administración & dosificación , Proteínas Klotho , Ratones , Modelos Animales , Sustancias Protectoras/administración & dosificaciónRESUMEN
This work reports the synthesis of a series of small-molecule-drug conjugates containing the αV ß3 -integrin ligand cyclo[DKP-RGD] or cyclo[DKP-isoDGR], a lysosomally cleavable Val-Ala (VA) linker or an "uncleavable" version devoid of this sequence, and monomethyl auristatinâ E (MMAE) or F (MMAF) as the cytotoxic agent. The conjugates were obtained via a straightforward synthetic scheme taking advantage of a copper-catalyzed azide-alkyne cycloaddition as the key step. The conjugates were tested for their binding affinity for the isolated αv ß3 receptor and were shown to retain nanomolar IC50 values, in the same range as those of the free ligands. The cytotoxic activity of the conjugates was evaluated in cell viability assays with αv ß3 integrin overexpressing human glioblastoma (U87) and human melanoma (M21) cells. The conjugates possess markedly lower cytotoxic activity than the free drugs, which is consistent with inefficient integrin-mediated internalization. In almost all cases the conjugates featuring isoDGR as integrin ligand exhibited higher potency than their RGD counterparts. In particular, the cyclo[DKP-isoDGR]-VA-MMAE conjugate has low nanomolar IC50 values in cell viability assays with both cancer cell lines tested (U87: 11.50±0.13â nm; M21: 6.94±0.09â nm) and is therefore a promising candidate for inâ vivo experiments.
Asunto(s)
Integrina alfaVbeta3/efectos de los fármacos , Oligopéptidos/síntesis química , Oligopéptidos/farmacología , Antineoplásicos/farmacología , Unión Competitiva , Línea Celular Tumoral , Reacción de Cicloadición , Evaluación Preclínica de Medicamentos , Humanos , Integrina alfaVbeta3/metabolismoRESUMEN
Mammalian pregnancy requires specific interactions between the conceptus and its mother that involve the endocrine system and adhesion molecules. The relation between adhesion molecules and their ligands at the fetal-maternal interface is crucial for developing a successful implantation. Progesterone (P4) and estrogen (E2) secreted by the porcine conceptus are required for the relation to be established. We investigated the expression of αvß3 integrin and its ligand, fibronectin (FN), at the placental interface, and E2 and P4 concentrations in both serum and maternal and fetal placental extracts during placentation in swine. Placental and serum samples of crossbred sows at 17, 30, 60, 70, and 114 days gestation and no pregnant uteri were used. The presence of αvß3 and FN were determined by immunohistochemistry, and E2 and P4 by chemiluminescence in homogenates of nonpregnant uterus (HoU), swine maternal placenta (HoPM), swine fetal placenta (HoPF) and serum. The expression of αvß3 and FN increased at the interface at 17, 30 and 60 days gestation. Immunostaining decreased by 70 days. Serum E2 levels peaked at 17 days, then decreased, then increased again near term. The highest concentration of P4 occurred in HoPF at 70 days gestation, then decreased coincident with a decline in integrin and FN expression at the placental interface. High P4 levels during swine gestation may regulate the expression of αvß3 integrin and FN at the placental interface for up to 70 days gestation. Other adhesion molecules and their ligands likely maintain the fetal-placental interface after 70 days.
Asunto(s)
Estrógenos/farmacología , Fibronectinas/metabolismo , Integrina alfaVbeta3/metabolismo , Placentación , Progesterona/farmacología , Animales , Estrógenos/sangre , Femenino , Fibronectinas/efectos de los fármacos , Inmunohistoquímica , Integrina alfaVbeta3/efectos de los fármacos , Embarazo , Progesterona/sangre , Estándares de Referencia , PorcinosRESUMEN
BACKGROUND: The resistance of melanoma cells to cisplatin restricts its clinical use. Therefore, the search for novel tumor inhibitors and effective combination treatments that sensitize tumor cells to this drug are still needed. We purified macrovipecetin, a novel heterodimeric C-type lectin, from Macrovipera lebetina snake venom and investigated its anti-tumoral effect on its own or combined with cisplatin, in human melanoma cells. METHODS: Biochemical characterization, in vitro cells assays such as viability, apoptosis, adhesion, migration, invasion, Western blotting and in silico analysis were used in this study. RESULTS: Macrovipecetin decreased melanoma cell viability 100 times more than cisplatin. Interestingly, when combined with the drug, macrovipecetin enhanced the sensitivity of SK-MEL-28 cells by augmenting their apoptosis through increased expression of the apoptosis inducing factor (AIF) and activation of ERK1/2, p38, AKT and NF-κB. Moreover, macrovipecetin alone or combined with cisplatin induced the expression of TRADD, p53, Bax, Bim and Bad and down-regulated the Bcl-2 expression and ROS levels in SK-MEL-28 cells. Interestingly, these treatments impaired SK-MEL-28 cell adhesion, migration and invasion through modulating the function and expression of αvß3 integrin along with regulating E-cadherin, vimentin, ß-catenin, c-Src and RhoA expression. In silico study suggested that only the α chain of macrovipecetin interacts with a region overlapping the RGD motif binding site on this integrin. CONCLUSIONS: We validated the antitumor effect of macrovipecetin when combined, or not, with cisplatin on SK-MEL-28 cells. GENERAL SIGNIFICANCE: The presented work proposes the potential use of macrovipecetin and cisplatin in combination as an effective anti-melanoma treatment.
Asunto(s)
Antineoplásicos/farmacología , Lectinas Tipo C/aislamiento & purificación , Melanoma/patología , Venenos de Víboras/química , Viperidae/metabolismo , Secuencia de Aminoácidos , Animales , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos Alquilantes/farmacología , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/biosíntesis , Proteínas Reguladoras de la Apoptosis/genética , Adhesión Celular/efectos de los fármacos , Moléculas de Adhesión Celular/biosíntesis , Moléculas de Adhesión Celular/genética , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Cisplatino/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Lectinas Tipo C/química , Modelos Moleculares , Simulación del Acoplamiento Molecular , Invasividad Neoplásica , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Conformación Proteica , Dominios y Motivos de Interacción de Proteínas , Mapeo de Interacción de Proteínas , Alineación de Secuencia , Homología de Secuencia de AminoácidoRESUMEN
In this study, we report the efficacy of RGD (arginine-glycine-aspartic acid) peptide-modified polylactic acid-co-glycolic acid (PLGA)-Chitosan nanoparticle (CSNP) for integrin αvß3 receptor targeted paclitaxel (PTX) delivery in lung cancer cells and its impact on normal cells. RGD peptide-modified chitosan was synthesized and then coated onto PTX-PLGA nanoparticles prepared by emulsion-solvent evaporation. PTX-PLGA-CSNP-RGD displayed favorable physicochemical properties for a targeted drug delivery system. The PTX-PLGA-CSNP-RGD system showed increased uptake via integrin receptor mediated endocytosis, triggered enhanced apoptosis, and induced G2/M cell cycle arrest and more overall cytotoxicity than its non-targeted counterpart in cancer cells. PTX-PLGA-CSNP-RGD showed less toxicity in lung fibroblasts than in cancer cells, may be attributed to low drug sensitivity, nevertheless the study invited close attention to their transient overexpression of integrin αvß3 and cautioned against corresponding uptake of toxic drugs, if any at all. Whereas, normal human bronchial epithelial (NHBE) cells with poor integrin αvß3 expression showed negligible toxicity to PTX-PLGA-CSNP-RGD, at equivalent drug concentrations used in cancer cells. Further, the nanoparticle demonstrated its capacity in targeted delivery of Cisplatin (CDDP), a drug having physicochemical properties different to PTX. Taken together, our study demonstrates that PLGA-CSNP-RGD is a promising nanoplatform for integrin targeted chemotherapeutic delivery to lung cancer.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Sistemas de Liberación de Medicamentos , Integrina alfaVbeta3/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Nanopartículas , Paclitaxel/administración & dosificación , Células A549/efectos de los fármacos , Antineoplásicos Fitogénicos/uso terapéutico , Línea Celular Tumoral , Quitosano , Sistemas de Liberación de Medicamentos/métodos , Humanos , Nanopartículas/administración & dosificación , Paclitaxel/uso terapéutico , Copolímero de Ácido Poliláctico-Ácido PoliglicólicoRESUMEN
BACKGROUND: Transforming growth factor-beta (TGF-ß) plays a central role in the pathogenesis of oral submucous fibrosis (OSF). Thrombin is a key player in tissue repair, inflammation, and fibrosis after injury. METHODS: Effects of thrombin on activated-TGF-ß1 levels, Smad3 phosphorylation, and connective tissue growth factor (CTGF/CCN2) synthesis in primary human buccal mucosal fibroblasts (BMFs) were assessed by enzyme-linked immunosorbent assay or Western blot analysis. RESULTS: Thrombin and protease-activated receptor-1 (PAR-1) agonist induced TGF-ß1 activation and Smad3 phosphorylation. Pretreatment with TGF-ß-neutralizing antibody completely inhibited thrombin-induced CCN2 synthesis. Neutralizing antibodies to integrin αv, ß1, αvß3, αvß5, and Rho-associated coiled-coil forming protein kinase (ROCK) inhibitor Y27632 completely blocked thrombin-induced TGF-ß1 activation, Smad3 phosphorylation, and CCN2 synthesis. Epigallocatechin-3-gallate (EGCG) dose-dependently inhibited thrombin-induced TGF-ß1 activation. CONCLUSION: Thrombin induces αvß1, αvß3, and αvß5 integrins-mediated TGF-ß1 activations via ROCK signaling. EGCG inhibits thrombin-induced CCN2 synthesis in BMFs by suppressing latent TGF-ß1 activation.
Asunto(s)
Integrina alfaVbeta3/metabolismo , Fibrosis de la Submucosa Bucal/patología , Receptores de Vitronectina/metabolismo , Trombina/farmacología , Factor de Crecimiento Transformador beta1/metabolismo , Western Blotting/métodos , Catequina/análogos & derivados , Catequina/metabolismo , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Mucosa Bucal/metabolismo , Mucosa Bucal/patología , Fibrosis de la Submucosa Bucal/metabolismo , Sensibilidad y Especificidad , Factor de Crecimiento Transformador beta1/efectos de los fármacosRESUMEN
UNLABELLED: Noninvasive, real-time, quantitative measurement of key biomarkers associated with cancer therapeutic interventions could provide a better understanding of cancer biology. We investigated in this study whether incorporating multiple molecular imaging approaches could be used to guide dasatinib anti-Src therapy and aid in the rational design of a combination therapy regimen. METHODS: Bioluminescence imaging, (18)F-FDG PET, integrin αvß3-targeted SPECT/CT, and vascular endothelial growth factor-targeted near-infrared fluorescence imaging were performed before and after dasatinib treatment in a tumor mouse model. RESULTS: There was no significant difference in the bioluminescence imaging signal or (18)F-FDG tumor uptake in dasatinib-treated tumors compared with the control tumors. However, the uptake of (99m)T-3PRGD2 (integrin αvß3-specific) and DyLight755-ranibizumab (vascular endothelial growth factor-specific) in the dasatinib-treated tumors was significantly lower than that in the control tumors. In vitro studies confirmed the antiangiogenic effects of dasatinib but indicated a lack of cytotoxicity. Dasatinib plus cytotoxic docetaxel elicited marked synergistic tumor growth inhibition in vivo. CONCLUSION: Visualization of post-Src inhibition tumor signatures through multiple imaging approaches facilitates sensitive and quantitative measurement of cancer biomarkers in vivo, thus aiding in the rational design of dasatinib combination therapy.
Asunto(s)
Antineoplásicos/uso terapéutico , Dasatinib/uso terapéutico , Genes src/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Animales , Quimioterapia Combinada , Fluorodesoxiglucosa F18/farmacocinética , Integrina alfaVbeta3/efectos de los fármacos , Luminiscencia , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Imagen Molecular , Ensayo de Unión Radioligante , Radiofármacos/farmacocinética , Ranibizumab/farmacocinética , Tomografía Computarizada de Emisión de Fotón Único , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacosRESUMEN
Type XIX collagen is a minor collagen associated with basement membranes. It was isolated for the first time in a human cDNA library from rhabdomyosarcoma and belongs to the FACITs family (Fibril Associated Collagens with Interrupted Triple Helices). Previously, we demonstrated that the NC1 domain of collagen XIX (NC1(XIX)) exerts anti-tumor properties on melanoma cells by inhibiting their migration and invasion. In the present work, we identified for the first time the integrin αvß3 as a receptor of NC1(XIX). Moreover, we demonstrated that NC1(XIX) inhibits the FAK/PI3K/Akt/mTOR pathway, by decreasing the phosphorylation and activity of the major proteins involved in this pathway. On the other hand, NC1(XIX) induced an increase of GSK3ß activity by decreasing its degree of phosphorylation. Treatments targeting this central signaling pathway in the development of melanoma are promising and new molecules should be developed. NC1(XIX) seems to have the potential for the design of new anti-cancer drugs.
Asunto(s)
Colágeno/metabolismo , Colágenos Asociados a Fibrillas/metabolismo , Quinasa 1 de Adhesión Focal/metabolismo , Integrina alfaVbeta3/metabolismo , Melanoma/enzimología , Fragmentos de Péptidos/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Neoplasias Cutáneas/enzimología , Serina-Treonina Quinasas TOR/metabolismo , Proteínas Quinasas Dependientes de 3-Fosfoinosítido/metabolismo , Antineoplásicos/farmacología , Línea Celular Tumoral , Colágeno/farmacología , Colágenos Asociados a Fibrillas/farmacología , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Melanoma/tratamiento farmacológico , Melanoma/patología , Terapia Molecular Dirigida , Fragmentos de Péptidos/farmacología , Fosforilación , Dominios Proteicos , Transducción de Señal/efectos de los fármacos , Neoplasias Cutáneas/tratamiento farmacológico , Neoplasias Cutáneas/patologíaRESUMEN
INTRODUCTION: This study evaluates the concentration and time-dependent effects of endodontic sealers' extracts (AH Plus [Dentsply DeTrey, Konstanz, Germany], GuttaFlow [Roeko, Colténe/Whaledent, Germany], Tubliseal [Kerr/Sybron, Romulus, MI], Sealapex [Kerr/Sybron, Romulus, MI], and RealSeal [SybronEndo, Orange, CA]) in the differentiation and function of both unstimulated and stimulated osteoclast precursors, simulating, respectively, immature/undifferentiated precursors and cells undergoing osteoclastogenesis. METHODS: The sealers were mixed according to the manufacturers' instructions, freshly extracted with culture medium (1.3 cm(2)/mL, 24 hours, 37°C, 5% CO2/air), and diluted (1:20, 1:100, 1:500, and 1:2500). Human peripheral blood mononuclear cells were used as osteoclast precursor cells. After overnight attachment, peripheral blood mononuclear cell cultures were exposed to the sealers' extracts during 21 days in the absence (unstimulated) or presence (stimulated) of recombinant macrophage colony-stimulating factor and receptor for the activation of nuclear factor-κB ligand. Cultures performed in the absence of the extracts were used as the control. Cultures were characterized for osteoclastic differentiation and function. RESULTS: Extracts caused mostly inhibitory effects on osteoclastic cells, both in unstimulated and stimulated conditions, which were reflected by a decrease in tartrate-resistant acid phosphatase activity, the presence of actin rings, vitronectin and calcitonin receptors, the calcium phosphate resorbing ability, and the expression of osteoclastic genes. Also, the extracts induced alterations in the relative contribution of some intracellular signaling pathways involved in osteoclastogenic events. The sealers differed in the dose- and time-dependent profile. An adaptive cell response was noticed for the inhibitory effects after long-term exposure. CONCLUSIONS: Endodontic sealers affect the osteoclastic differentiation and activity, which is followed by an adaptive cell response. Our results suggest that the deleterious effect in the bone periapical tissues observed with the root canal sealers might involve, at least partially, a direct effect on the osteoclastic cells.
Asunto(s)
Osteoclastos/efectos de los fármacos , Materiales de Obturación del Conducto Radicular/farmacología , Fosfatasa Ácida/efectos de los fármacos , Actinas/efectos de los fármacos , Adulto , Hidróxido de Calcio/química , Hidróxido de Calcio/farmacología , Fosfatos de Calcio/química , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Resinas Compuestas/química , Resinas Compuestas/farmacología , Dimetilpolisiloxanos/química , Dimetilpolisiloxanos/farmacología , Relación Dosis-Respuesta a Droga , Combinación de Medicamentos , Resinas Epoxi/química , Resinas Epoxi/farmacología , Gutapercha/química , Gutapercha/farmacología , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Isoenzimas/efectos de los fármacos , Leucocitos Mononucleares/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Factor Estimulante de Colonias de Macrófagos/farmacología , Masculino , FN-kappa B/efectos de los fármacos , Proteína Quinasa C/efectos de los fármacos , Ligando RANK/farmacología , Receptores de Calcitonina/efectos de los fármacos , Materiales de Obturación del Conducto Radicular/química , Salicilatos/química , Salicilatos/farmacología , Transducción de Señal/efectos de los fármacos , Fosfatasa Ácida Tartratorresistente , Factores de Tiempo , Cemento de Óxido de Zinc-Eugenol/química , Cemento de Óxido de Zinc-Eugenol/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/efectos de los fármacosRESUMEN
A small library of integrin ligand-paclitaxel conjugates 10-13 was synthesized with the aim of using the tumor-homing cyclo[DKP-RGD] peptidomimetics for site-directed delivery of the cytotoxic drug. All the paclitaxel-RGD constructs 10-13 inhibited biotinylated vitronectin binding to the purified αVß3 integrin receptor at low nanomolar concentration and showed in vitro cytotoxic activity against a panel of human tumor cell lines similar to that of paclitaxel. Among the cell lines, the cisplatin-resistant IGROV-1/Pt1 cells expressed high levels of integrin αVß3, making them attractive to be tested in in vivo models. cyclo[DKP-f3-RGD]-PTX 11 displayed sufficient stability in physiological solution and in both human and murine plasma to be a good candidate for in vivo testing. In tumor-targeting experiments against the IGROV-1/Pt1 human ovarian carcinoma xenotransplanted in nude mice, compound 11 exhibited a superior activity compared with paclitaxel, despite the lower (about half) molar dosage used.
Asunto(s)
Antineoplásicos Fitogénicos/química , Integrina alfaVbeta3/efectos de los fármacos , Oligopéptidos/química , Paclitaxel/química , Peptidomiméticos , Animales , Antineoplásicos Fitogénicos/farmacología , Línea Celular Tumoral , Femenino , Humanos , Inmunohistoquímica , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Masculino , Oligopéptidos/farmacología , Paclitaxel/farmacología , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
BACKGROUND: Integrin αvß3 plays an important role in the regulation of cell proliferation and neoangiogenesis. We found mesangial de novo expression of integrin αvß3 in mesangioproliferative glomerulonephritis (MesGN). The aim of the study was to clarify if blockade of αvß3 integrin with the specific αvß3-blocking cyclic peptide RGDdFV (cRGD) has beneficial effects on the course of this disease. METHODS: Habu snake venom (Habu) GN was induced in male C57BL/6 mice 1 week after uninephrectomy (6 mg Habu toxin/kg body weight intravenously). After 24 h, nephritic animals received αvß3-inhibitory cRGD or cRAD control peptides for 3 or 7 days, respectively. The kidneys were investigated using morphometry, immunohistochemistry and TaqMan polymerase chain reaction. RESULTS: At Day 3, serum creatinine and albuminuria were lower after cRGD compared to cRAD treatment. At Day 3, glomerulosclerosis index, percentage of glomerular injury, mesangial cell (MC) number and volume density of mesangial matrix were significantly lower (P < 0.05) in cRGD-treated mice than in cRAD-treated controls. At Day 7, only a mild effect of cRGD on mesangial matrix expansion and fibronectin messenger RNA was still detectable (P < 0.05). Complementary in vitro studies in MCs revealed that inhibition of αvß3 by cRGD-blocked adhesion, reduced proliferation and increased apoptosis of MCs. CONCLUSION: Habu GN inhibition of integrin αvß3 by cRGD partly ameliorates early injury but has no or only mild effects on late glomerular lesions.
Asunto(s)
Glomerulonefritis/tratamiento farmacológico , Integrina alfaVbeta3/antagonistas & inhibidores , Integrina alfaVbeta3/efectos de los fármacos , Oligopéptidos/farmacología , Oligopéptidos/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Creatinina/sangre , Venenos de Crotálidos/efectos adversos , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Glomerulonefritis/etiología , Glomerulonefritis/patología , Técnicas In Vitro , Masculino , Células Mesangiales/efectos de los fármacos , Células Mesangiales/patología , Ratones , Ratones Endogámicos C57BL , Factores de TiempoRESUMEN
Parathyroid hormone (PTH) is widely accepted as an anabolic agent when administered intermittently. Here, we explored the influence of intermittent PTH(1-34) on the expression of local factors by human periodontal ligament (PDL) cells that modify osteoclast biology. This approach aimed at a further elucidation of the role of the hormone and of PDL cells in the regulation of periodontal tissue homeostasis and of repair processes. In a co-culture model of mature PDL cells and RAW 264.7 cells, intermittent PTH(1-34) induced an increased gene expression for tartrate-resistant acid phosphatase (+84%), cathepsin K (+56%), and vitronectin-receptor (+56%); and an enhanced resorptive activity of differentiated osteoclasts (+154%). These findings were correlated with a reduction of the osteoprotegerin (OPG)/receptor activator of nuclear factor kappaB ligand (RANKL) ratio in the presence of PTH(1-34; -44%). Similar results were obtained when RAW cells were cultured with the conditioned medium of PTH(1-34)-stimulated PDL cells. In contrast, when less mature PDL cells were co-cultured with RAW cells, PTH(1-34) induced an inhibition of osteoclastic differentiation (TRAP, -35%; cathepsin K, -28%; vitronectin-receptor, -35%), a reduction of the resorbed substrate area (-77%) and an increase of the OPG/RANKL ratio (+11%). The conditioned medium of PTH(1-34)-pretreated less mature PDL cells led to a down-regulation of the number and activity of multinucleated cells. These data indicate that intermittent PTH(1-34) modifies the expression of membrane-bound and secreted factors by PDL cells which then in turn alter osteoclast biology. The PDL cell response to PTH(1-34) is specific in terms of cell maturation and the mechanism involved.
Asunto(s)
Osteoclastos/efectos de los fármacos , Osteoprotegerina/efectos de los fármacos , Hormona Paratiroidea/farmacología , Ligamento Periodontal/efectos de los fármacos , Ligando RANK/efectos de los fármacos , Fosfatasa Ácida/efectos de los fármacos , Adolescente , Resorción Ósea/fisiopatología , Catepsina K/efectos de los fármacos , Técnicas de Cultivo de Célula , Diferenciación Celular/efectos de los fármacos , Línea Celular , Células Cultivadas , Niño , Técnicas de Cocultivo , Medios de Cultivo Condicionados , Regulación hacia Abajo , Homeostasis/efectos de los fármacos , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Isoenzimas/efectos de los fármacos , Osteoclastos/fisiología , Ligamento Periodontal/citología , Regeneración/efectos de los fármacos , Fosfatasa Ácida TartratorresistenteRESUMEN
Interstitial lung disease (ILD) produces disruption of alveolar walls with loss of functionality and scar tissue accumulation. Asbestosis is the ILD produced by the inhalation of asbestos fibers. This study attempts to elucidate the role of lung epithelial cells in the generation of asbestos-induced ILD. When exposed to crocidolite LA-4 cells had a decrease in viability and an increase in the release of lactate dehydrogenase (LDH) and 6-keto PGF(1alpha), a PGI(2) metabolite. PGI(2) release was mediated by cyclooxygenase-2 (COX-2) and vitronectin receptor (VNR). When LA-4 cells were treated with VNR inhibitors, either RGD (Arg-Gly-Asp) peptide or VNR blocking antibody, a statistically significant decrease in PGI(2) metabolite production was observed, but crocidolite-induced cytotoxicity was not prevented. These findings propose that crocidolite is coated by an RGD protein and binds VNR-inducing COX-2 expression and PGI(2) release. Moreover, when LA-4 cells were exposed to crocidolite in the presence of reduced serum culture media, PGI(2) production was prevented, and when bronchoalveolar lavage fluid (BALF) was added, PGI(2) production was rescued. Cytotoxicity did not occur, either in reduced serum culture media or when BALF was added. In conclusion, crocidolite requires the presence of an RGD protein coating the fibers to induce inflammation (PGI(2) production) and crocidolite alone cannot induce cytotoxicity in lung cells.
Asunto(s)
Asbesto Crocidolita/toxicidad , Ciclooxigenasa 2/metabolismo , Células Epiteliales/efectos de los fármacos , Epoprostenol/metabolismo , Integrina alfaVbeta3/efectos de los fármacos , Pulmón/efectos de los fármacos , 6-Cetoprostaglandina F1 alfa/metabolismo , Animales , Líquido del Lavado Bronquioalveolar/química , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células Epiteliales/enzimología , Células Epiteliales/patología , Integrina alfaVbeta3/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Pulmón/enzimología , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Oligopéptidos/toxicidad , Transducción de Señal/efectos de los fármacosRESUMEN
There is mounting evidence that tumors are initiated by a rare subset of cells called cancer stem cells (CSCs). CSCs are generally quiescent, self-renew, form tumors at low numbers, and give rise to the heterogeneous cell types found within a tumor. CSCs isolated from multiple tumor types differentiate both in vivo and in vitro when cultured in serum, yet the factors responsible for their differentiation have not yet been identified. Here we show that vitronectin is the component of human serum driving stem cell differentiation through an integrin alpha V beta 3-dependent mechanism. CSCs cultured on vitronectin result in downregulation of stem cell genes, modulation of differentiation markers, and loss of beta-catenin nuclear localization. Blocking integrin alpha V beta 3 inhibits differentiation and subsequently tumor formation. Thus, CSCs must be engaged by one or more extracellular signals to differentiate and initiate tumor formation, defining a new axis for future novel therapies aimed at both the extrinsic and intracellular pathways.
Asunto(s)
Transformación Celular Neoplásica/metabolismo , Neoplasias/metabolismo , Células Madre Neoplásicas/metabolismo , Vitronectina/metabolismo , Animales , Biomarcadores/análisis , Biomarcadores/metabolismo , Proteínas Sanguíneas/metabolismo , Proteínas Sanguíneas/farmacología , Neoplasias de la Mama/inducido químicamente , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/fisiopatología , Carcinoma/inducido químicamente , Carcinoma/metabolismo , Carcinoma/fisiopatología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Línea Celular Tumoral , Transformación Celular Neoplásica/efectos de los fármacos , Cromatografía Liquida , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Integrina alfaVbeta3/metabolismo , Masculino , Espectrometría de Masas , Ratones , Neoplasias/inducido químicamente , Neoplasias/fisiopatología , Células Madre Neoplásicas/efectos de los fármacos , Señales de Localización Nuclear/efectos de los fármacos , Señales de Localización Nuclear/metabolismo , Neoplasias de la Próstata/inducido químicamente , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/fisiopatología , Vitronectina/farmacología , beta Catenina/efectos de los fármacos , beta Catenina/metabolismoRESUMEN
The purpose of this study was to assess whether an alternative treatment approach that targets angiogenesis, delivered through ligand-targeted nanotherapy, would ameliorate inflammatory arthritis. Arthritis was induced using the K/BxN mouse model of inflammatory arthritis. After arthritis was clearly established, mice received three consecutive daily doses of alpha(v)beta(3)-targeted fumagillin nanoparticles. Control groups received no treatment or alpha(v)beta(3)-targeted nanoparticles without drugs. Disease score and paw thickness were measured daily. Mice that received alpha(v)beta(3)-targeted fumagillin nanoparticles showed a significantly lower disease activity score (mean score of 1.4+/-0.4; P<0.001) and change in ankle thickness (mean increase of 0.17+/-0.05 mm; P<0.001) 7 d after arthritis induction, whereas the group that received alpha(v)beta(3)-targeted nanoparticles without drugs exhibited a mean arthritic score of 9.0 +/- 0.3 and mean change in ankle thickness of 1.01 +/- 0.09 mm. Meanwhile, the group that received no treatment showed a mean arthritic score of 9.8 +/- 0.5 and mean change in ankle thickness of 1.05 +/- 0.10 mm. Synovial tissues from animals treated with targeted fumagillin nanoparticles also showed significant decrease in inflammation and angiogenesis and preserved proteoglycan integrity. Ligand-targeted nanotherapy to deliver antiangiogenic agents may represent an effective way to treat inflammatory arthritis.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Ciclohexanos/uso terapéutico , Ácidos Grasos Insaturados/uso terapéutico , Integrina alfaVbeta3/efectos de los fármacos , Animales , Artritis Experimental/patología , Ciclohexanos/farmacología , Sistemas de Liberación de Medicamentos/métodos , Ácidos Grasos Insaturados/farmacología , Inflamación/tratamiento farmacológico , Ratones , Nanopartículas/uso terapéutico , Neovascularización Patológica/tratamiento farmacológico , Sesquiterpenos/farmacología , Sesquiterpenos/uso terapéutico , Resultado del TratamientoRESUMEN
As large amounts of porcine cartilage are discarded as waste in daily life, it is necessary to find new uses for them. We extracted polysaccharide from cartilage and performed in vitro and in vivo experiments in cancer cells. A mouse breast-cancer pulmonary metastasis model was set up, and we tried to determine the mechanism of the inhibition of metastasis by cartilage PS (polysaccharide). Effects on tumour size and the progression of metastasis indicated that cartilage PS can obviously inhibit metastasis in breast-cancer cells. The levels of LNR1 (laminin receptor 1), alphavbeta3 integrin and MMP-9 (matrix metalloproteinase-9) in mice treated or not with cartilage PS showed significant differences. Cartilage PS inhibited the growth of MCF-7 human breast adenocarcinoma cells, but had little effect on normal cells. Cartilage PS can inhibit the activity of the MMP-2 and the MMP-9 by decreasing the levels of LNR1 and alphavbeta3 integrin to inhibit metastasis further. In summary, we conclude that cartilage PS can act as a specific anti-metastatic agent in breast-cancer cells.
Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Cartílago/química , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/secundario , Metástasis de la Neoplasia/tratamiento farmacológico , Polisacáridos/farmacología , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Humanos , Integrina alfaVbeta3/efectos de los fármacos , Integrina alfaVbeta3/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidores de la Metaloproteinasa de la Matriz , Ratones , Trasplante de Neoplasias , Polisacáridos/aislamiento & purificación , Receptores de Laminina/efectos de los fármacos , Receptores de Laminina/metabolismo , Proteínas Ribosómicas , PorcinosRESUMEN
OBJECTIVE: To compare the effects of letrozole and clomiphene citrate on the expression of HOXA10 and integrin alpha(v)beta(3) in the uterine epithelium in rats. DESIGN: Controlled prospective study. SETTING: Teaching hospital and university research laboratory. ANIMAL(S): Sixty sexually mature female Wistar-Albino rats that were 20 weeks of age. INTERVENTION(S): Letrozole, 5 mg/kg of body weight daily (20 rats); clomiphene citrate, 100 microg/kg daily (20 rats); and saline solution, 2 mL/day (20 rats). After 2 days, rats were killed and hysterectomized. MAIN OUTCOME MEASURE(S): Expression of integrin alpha(v)beta(3) and HOXA10. RESULT(S): The expression of integrin alpha(v)beta(3) in the clomiphene citrate group was statistically significantly lower than in the letrozole and saline solution groups. The expression of HOXA10 was statistically significantly higher in the saline solution group than in the letrozole group, and the letrozole group showed a statistically significantly higher expression of HOXA10 compared with the clomiphene citrate group. CONCLUSION(S): In rats, letrozole affects the expression of HOXA10 in uterine epithelium but has no effect on the expression of integrin alpha(v)beta(3), which suggests that clomiphene suppresses endometrial receptivity more than letrozole. In the future, letrozole may be an appropriate drug for ovulation induction.