RESUMEN
Cisplatin, the first platinum compound approved for cancer treatment, is widely used in the treatment of various cancers including hepatocellular carcinoma (HCC). HCC incidence rates rise globally. Epithelial mesenchymal transition (EMT) is implicated in cancer invasion and metastasis, which are associated with increased mortality. Cisplatin dose might influence cancer invasion and metastatic behavior of the cells. The aim of the study was to investigate the effect of low-dose cisplatin treatment on EMT- related changes in HepG2 cells. Following treatment with 4 µM cisplatin, HepG2 cells were evaluated morphologically. Gene expression of E-cadherin, Vimentin, Snail1 was assessed by quantitative PCR. Immunofluorescence analyses of NA-K ATPase were performed. Although the low-dose cisplatin treated cells exhibited a more stretched morphology, no statistical difference was detected in gene expression of E-cadherin, Vimentin, Snail1 and immunofluorescence of NA-K ATPase. Findings on low-dose cisplatin effects in HepG2 might contribute to the knowledge of antineoplastic inefficacy by further understanding the molecular mechanisms of drug action.
El cisplatino, el primer compuesto de platino aprobado para el tratamiento del cáncer, es ampliamente utilizado en el tratamiento de varios tipos de cáncer, incluido el carcinoma hepatocelular (CHC). Las tasas de incidencia de CHC aumentan a nivel mundial. La transición mesenquimal epitelial (EMT) está implicada en la invasión del cáncer y la metástasis, que se asocian con un aumento de la mortalidad. La dosis de cisplatino podría influir en la invasión del cáncer y el comportamiento metastásico de las células. El objetivo del estudio fue investigar el efecto del tratamiento con dosis bajas de cisplatino en los cambios relacionados con la EMT en las células HepG2. Tras el tratamiento con cisplatino de 4 µM, se evaluaron morfológicamente las células HepG2. La expresión génica de E-cadherina, vimentina, caracol1 se evaluó mediante PCR cuantitativa. Se realizaron análisis de inmunofluorescencia de NA-K ATPasa . Aunque las células tratadas con cisplatino en dosis bajas exhibieron una morfología más estirada, no se detectaron diferencias estadísticas en la expresión génica de E-cadherina, vimentina, Snail1 e inmunofluorescencia de NA-K ATPasa. Los hallazgos sobre los efectos del cisplatino en dosis bajas en HepG2 podrían contribuir al conocimiento de la ineficacia antineoplásica al comprender mejor los mecanismos moleculares de la acción del fármaco.
Asunto(s)
Humanos , Cisplatino/administración & dosificación , Antineoplásicos/administración & dosificación , Vimentina/efectos de los fármacos , Vimentina/genética , Vimentina/metabolismo , Cadherinas/efectos de los fármacos , Cadherinas/genética , Cadherinas/metabolismo , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Microscopía Confocal , Células Hep G2 , Transición Epitelial-Mesenquimal , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de Transcripción de la Familia Snail/efectos de los fármacos , Factores de Transcripción de la Familia Snail/genética , Factores de Transcripción de la Familia Snail/metabolismo , Invasividad NeoplásicaRESUMEN
SUMMARY: Carnosine is known as a natural dipeptide, which inhibits the proliferation of tumor cells throughout its action on mitochondrial respiration and cell glycolysis. However, not much is known about its effects on the metabolism of healthy cells. We explored the effects of Karnozin EXTRA® capsule with different concentrations of L-carnosine, on the cell viability and the expressions of intermediate filament vimentin (VIM) and superoxide dismutase (SOD2) in normal fibroblasts BHK-21/C13. Furthermore, we investigated its action on the energy production of these cells. Cell viability was quantified by the MTT assay. The Clark oxygen electrode (Oxygraph, Hansatech Instruments, England) was used to measure the "intact cell respiration rate", state 3 of ADP-stimulated oxidation, maximum oxidation capacity and the activities of complexes I, II and IV. Results showed that Karnozin EXTRA® capsule in concentrations of 2 and 5 mM of L-carnosine did not induce toxic effects and morphological changes in treated cells. Our data revealed a dose-dependent immunofluorescent signal amplification of VIM and SOD2 in the BHK-21/C13 cell line. This supplement substantially increased the recorded mitochondrial respiration rates in the examined cell line. Due to the stimulation of mitochondrial energy production in normal fibroblasts, our results suggested that Karnozin EXTRA® is a potentially protective dietary supplement in the prevention of diseases with altered mitochondrial function.
RESUMEN: La carnosina se conoce como dipéptido natural, que inhibe la proliferación de células tumorales a través de su acción sobre la respiración mitocondrial y la glucólisis celular. Sin embargo, no se sabe mucho de sus efectos sobre el metabolismo de las células sanas. Exploramos los efectos de la cápsula Karnozin EXTRA® con diferentes concentraciones de L-carnosina, sobre la viabilidad celular y las expresiones de vimentina de filamento intermedio (VIM) y superóxido dismutasa (SOD2) en fibroblastos normales BHK-21 / C13. Además, estudiamos su acción sobre la producción de energía de estas células. La viabilidad celular se cuantificó mediante el ensayo MTT. Se utilizó el electrodo de oxígeno Clark (Oxygraph, Hansatech Instruments, Inglaterra) para medir la "tasa de respiración de células intactas", el estado 3 de oxidación estimulada por ADP, la capacidad máxima de oxidación y las actividades de los complejos I, II y IV. Los resultados mostraron que la cápsula de Karnozin EXTRA® en concentraciones de 2 y 5 mM de L- carnosina no indujo efectos tóxicos ni cambios morfológicos en las células tratadas. Nuestros datos revelaron una amplificación de señal inmunofluorescente dependiente de la dosis de VIM y SOD2 en la línea celular BHK-21 / C13. Este suplemento aumentó sustancialmente las tasas de respiración mitocondrial registradas en la línea celular examinada. Debido a la estimulación de la producción de energía mitocondrial en fibroblastos normales, nuestros resultados sugirieron que Karnozin EXTRA® es un suplemento dietético potencialmente protector en la prevención de enfermedades con función mitocondrial alterada.
Asunto(s)
Animales , Carnosina/farmacología , Fibroblastos/efectos de los fármacos , Riñón/citología , Superóxido Dismutasa/efectos de los fármacos , Vimentina/efectos de los fármacos , Bioensayo , Supervivencia Celular/efectos de los fármacos , Técnica del Anticuerpo Fluorescente , Cricetinae , Técnicas de Cultivo de Célula , Metabolismo EnergéticoRESUMEN
Hepatocellular carcinoma (HCC) is one of the most burdened tumors worldwide, with a complex and multifactorial pathogenesis. Current treatment approaches involve different molecular targets. Phytochemicals have shown considerable promise in the prevention and treatment of HCC. We investigated the efficacy of two natural components, 1,8 cineole (Cin) and ellagic acid (EA), against diethylnitrosamine/2-acetylaminofluorene (DEN/2-AAF) induced HCC in rats. DEN/2-AAF showed deterioration of hepatic cells with an impaired functional capacity of the liver. In addition, the levels of tumor markers including alpha-fetoprotein, arginase-1, alpha-L-fucosidase, and ferritin were significantly increased, whereas the hepatic miR-122 level was significantly decreased in induced-HCC rats. Interestingly, treatment with Cin (100mg/kg) and EA (60mg/kg) powerfully restored these biochemical alterations. Moreover, Cin and EA treatment exhibited significant downregulation in transforming growth factor beta-1 (TGF-ß1), Fascin-1 (FSCN1), vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), and epithelial-mesenchymal transition (EMT) key marker, vimentin, along with a restoration of histopathological findings compared to HCC group. Such effects were comparable to Doxorubicin (DOX) (2mg/kg); however, a little additive effect was evident through combining these phytochemicals with DOX. Altogether, this study highlighted 1,8 cineole and ellagic acid for the first time as promising phytochemicals for the treatment of hepatocarcinogenesis via regulating multiple targets.
Asunto(s)
Antineoplásicos/farmacología , Carcinoma Hepatocelular , Ácido Elágico , Eucaliptol , Fitoquímicos/farmacología , Animales , Carcinoma Hepatocelular/inducido químicamente , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Proteínas Portadoras/efectos de los fármacos , Proteínas Portadoras/metabolismo , Modelos Animales de Enfermedad , Ácido Elágico/administración & dosificación , Ácido Elágico/farmacología , Eucaliptol/administración & dosificación , Eucaliptol/farmacología , Humanos , Neoplasias Hepáticas/inducido químicamente , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Metaloproteinasa 9 de la Matriz/efectos de los fármacos , Metaloproteinasa 9 de la Matriz/metabolismo , MicroARNs/efectos de los fármacos , MicroARNs/metabolismo , Proteínas de Microfilamentos/efectos de los fármacos , Proteínas de Microfilamentos/metabolismo , Ratas , Factor de Crecimiento Transformador beta1/efectos de los fármacos , Factor de Crecimiento Transformador beta1/metabolismo , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/metabolismo , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
The incidence of colorectal cancer (CRC) is reported to be increasing nowadays, with a large proportion of newly diagnosed CRC patients being affected by metastasis. Epithelial-mesenchymal transition (EMT) is an important event in the development of metastasis of CRC. In this study, we investigated whether the anticancer drug bevacizumab and anexelekto inhibitor, TP-0903, regulate EMT of colon cancer cells induced by transforming growth factor-beta 1 (TGF-ß1). Using quantitative real-time PCR and western blot analysis, we found that bevacizumab and TP-0903 decreased the expression levels of fibronectin, alpha-smooth muscle actin, and vimentin, whereas they restored E-cadherin expression in TGF-ß1-exposed SW480 and HCT116 cells. In addition, we elucidated that bevacizumab and TP-0903 inhibited the migration and invasion of TGF-ß1-exposed colon cancer cells using scratched wound healing, transwell migration, and Matrigel-coated invasion assays. Finally, we discovered that bevacizumab and TP-0903 inactivated the Smad 2/3 signaling pathway in TGF-ß1-exposed SW480 and HCT116 cells. Therefore, we suggest that treatment of bevacizumab and TP-0903 inhibits TGF-ß1-induced EMT of colon cancer cells through inactivation of the Smad 2/3 signaling pathway.
Asunto(s)
Antineoplásicos Inmunológicos/farmacología , Bevacizumab/farmacología , Neoplasias del Colon/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Pirimidinas/farmacología , Sulfonamidas/farmacología , Factor de Crecimiento Transformador beta1/efectos de los fármacos , Actinas/efectos de los fármacos , Antineoplásicos Inmunológicos/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Bevacizumab/administración & dosificación , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Fibronectinas/efectos de los fármacos , Humanos , Proteínas Proto-Oncogénicas/antagonistas & inhibidores , Pirimidinas/administración & dosificación , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Sulfonamidas/administración & dosificación , Vimentina/efectos de los fármacos , Tirosina Quinasa del Receptor AxlRESUMEN
Vimentin is a cytoskeletal intermediate filament protein that plays pivotal roles in tumor initiation, progression, and metastasis, and its overexpression in aggressive cancers predicted poor prognosis. Herein described is a highly effective antitumor and antimetastatic metal complex [PtII(C^N^N)(NHC2Bu)]PF6 (Pt1a; HC^N^N = 6-phenyl-2,2'-bipyridine; NHC= N-heterocyclic carbene) that engages vimentin via noncovalent binding interactions with a distinct orthogonal structural scaffold. Pt1a displays vimentin-binding affinity with a dissociation constant of 1.06 µM from surface plasmon resonance measurements and fits into a pocket between the coiled coils of the rod domain of vimentin with multiple hydrophobic interactions. It engages vimentin in cellulo, disrupts vimentin cytoskeleton, reduces vimentin expression in tumors, suppresses xenograft growth and metastasis in different mouse models, and is well tolerated, attributable to biotransformation to less toxic and renal-clearable platinum(II) species. Our studies uncovered the practical therapeutic potential of platinum(II)âNHC complexes as effective targeted chemotherapy for combating metastatic and cisplatin-resistant cancers.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Compuestos Organoplatinos/uso terapéutico , Vimentina/efectos de los fármacos , Animales , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Femenino , Células HCT116 , Humanos , Neoplasias Pulmonares/secundario , Ratones , Ratones Desnudos , Simulación de Dinámica Molecular , Compuestos Organoplatinos/metabolismo , Compuestos Organoplatinos/farmacología , Ratas , Vimentina/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Stachydrine is a bioactive alkaloid that has been found to exert tumor-suppressive potential. However, the effect of stachydrine on hepatocellular carcinoma (HCC) has not been previously investigated. In the present study, we investigated the effect of transforming growth factor-ß1 (TGF-ß1)-induced epithelial-mesenchymal transition (EMT) in HepG2 cells. Our results showed that stachydrine significantly suppressed TGF-ß1-induced HepG2 cell migration and invasion in a dose-dependent manner. Stachydrine prevented TGF-ß1-induced EMT in HepG2 cells, as proved by the increased expression level of E-cadherin and decreased expression levels of N-cadherin and vimentin. In addition, stachydrine attenuated TGF-ß1-induced upregulation of TGF-ß receptor I (TßRI) in both protein and mRNA levels. Further mechanism investigations proved that stachydrine prevented TGF-ß1-induced activation of Smad2/3 and phosphoinositol-3-kinase (PI3K)/Akt/mTOR signaling pathways in HepG2 cells. In conclusion, these findings demonstrated that stachydrine prevented TGF-ß1-induced EMT in HCC cells through Smad2/3 and PI3K/Akt/mTOR signaling pathways. Thus, stachydrine might be a potential therapeutic agent for the treatment of HCC.
Asunto(s)
Carcinoma Hepatocelular/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Neoplasias Hepáticas/patología , Prolina/análogos & derivados , Factor de Crecimiento Transformador beta1/efectos de los fármacos , Cadherinas/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Supervivencia Celular , Relación Dosis-Respuesta a Droga , Células Hep G2 , Humanos , Fosfatidilinositol 3-Quinasas , Prolina/farmacología , Transducción de Señal/efectos de los fármacos , Proteína Smad2/efectos de los fármacos , Serina-Treonina Quinasas TOR/efectos de los fármacos , Vimentina/efectos de los fármacosRESUMEN
Alkaptonuria (AKU) is an ultra-rare disease caused by the deficient activity of homogentisate 1,2-dioxygenase enzyme, leading the accumulation of homogentisic acid (HGA) in connective tissues implicating the formation of a black pigmentation called "ochronosis." Although AKU is a multisystemic disease, the most affected tissue is the articular cartilage, which during the pathology appears to be highly damaged. In this study, a model of alkaptonuric chondrocytes and cartilage was realized to investigate the role of HGA in the alteration of the extracellular matrix (ECM). The AKU tissues lost its architecture composed of collagen, proteoglycans, and all the proteins that characterize the ECM. The cause of this alteration in AKU cartilage is attributed to a degeneration of the cytoskeletal network in chondrocytes caused by the accumulation of HGA. The three cytoskeletal proteins, actin, vimentin, and tubulin, were analyzed and a modification in their amount and disposition in AKU chondrocytes model was identified. Cytoskeleton is involved in many fundamental cellular processes; therefore, the aberration in this complex network is involved in the manifestation of AKU disease.
Asunto(s)
Cartílago Articular/efectos de los fármacos , Condrocitos/efectos de los fármacos , Citoesqueleto/efectos de los fármacos , Matriz Extracelular/efectos de los fármacos , Ácido Homogentísico/farmacología , Actinas/efectos de los fármacos , Actinas/metabolismo , Alcaptonuria/metabolismo , Cartílago Articular/metabolismo , Condrocitos/metabolismo , Citoesqueleto/metabolismo , Matriz Extracelular/metabolismo , Humanos , Microtúbulos/efectos de los fármacos , Microtúbulos/metabolismo , Ocronosis/tratamiento farmacológico , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
Clozapine (CLZ) represents an effective treatment for resistant schizophrenia. However, myocarditis, recently reported in about 66% of the psychiatric patients treated with CLZ, has raised concerns about its safety. ß-blocking agents have shown to be helpful in the management of myocarditis. Moreover, Vimentin (VIM) and Connexin-43 (CX43) are important structural proteins play key roles in cytoskeletal functions and cellular communication and have complex implications in pathophysiology. The present work aimed to study the mechanisms behind the protective effect of propranolol (PRO) against CLZ-induced myocarditis and the possible involvement of VIM and CX43. The effect of PRO (5 and 10 mg/kg, oral) on the myocarditis induced by CLZ (25 mg/kg/d, i. p.) treatment for 21 days in rats, was assessed biochemically, and immunohistochemically. CLZ treatment increased the serum levels of cardiac injury (CK-MP, LDH and cTn-I) and cardiac levels of oxidative stress (TBARS and NO) markers, proinflammatory cytokines (IL-1ß and TNF-α), and mRNA expression of VIM and CX43 with decreased the antioxidant defenses (GSH and GSH-Px). Immunohistochemical study showed increased cardiac expression of VIM, CX43 and caspase-3 proteins. Coadministration of PRO with CLZ, dose-dependently decreased the biochemical and immunohistochemical hallmarks of CLZ-induced myocardial injury and significantly decreased mRNA expression of VIM and CX43. Taken together, our results demonstrate that the cardioprotective effects of PRO on CLZ-induced myocarditis are related in addition to its ß-blocking activity to protection of myocardial VIM and CX43 proteins through antagonizing the CLZ-induced oxidative stress and inflammatory response, and preventing cell apoptosis.
Asunto(s)
Antagonistas Adrenérgicos beta/farmacología , Cardiotónicos/farmacología , Conexina 43/metabolismo , Miocarditis/prevención & control , Propranolol/farmacología , Vimentina/metabolismo , Antagonistas Adrenérgicos beta/uso terapéutico , Animales , Antipsicóticos/toxicidad , Apoptosis/efectos de los fármacos , Cardiotónicos/uso terapéutico , Caspasa 3/metabolismo , Clozapina/toxicidad , Conexina 43/efectos de los fármacos , Conexina 43/genética , Creatina Quinasa/sangre , Citocinas/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , L-Lactato Deshidrogenasa/sangre , Masculino , Miocarditis/inducido químicamente , Miocarditis/metabolismo , Miocardio/metabolismo , Miocardio/patología , Estrés Oxidativo/efectos de los fármacos , Propranolol/uso terapéutico , Ratas Wistar , Troponina I/sangre , Vimentina/efectos de los fármacos , Vimentina/genéticaRESUMEN
BACKGROUND: Aberrant activation of epidermal growth factor receptor (EGFR) signaling pathway is associated with the pathogenesis of pulmonary hypertension (PH). However, the effect of icotinib, a first generation of EGFR tyrosine kinase inhibitor (EGFR-TKI), on PH remains to be elucidated. METHODS: PH rat model was established by a single intraperitoneal injection of monocrotaline (MCT, 60 mg/kg). Icotinib (15, 30, and 60 mg/kg/day) was administered by oral gavage from the day of MCT injection. After 4 weeks, hemodynamic parameters and histological changes of the pulmonary arterial vessels were assessed, and the phenotypic switching of pulmonary arterial smooth muscle cells (PASMCs) was determined in vivo. Moreover, the effects of icotinib (10 µM) on epidermal growth factor (EGF, 50 ng/ml)-stimulated proliferation, migration, and phenotypic switching of human PASMCs were explored in vitro. RESULTS: Icotinib significantly reduced the right ventricular systolic pressure and right ventricle hypertrophy index in rats with MCT-induced PH. Moreover, icotinib improved MCT-induced pulmonary vascular remodeling. The expression of contractile marker (smooth muscle 22 alpha (SM22α)) and synthetic markers (osteopontin (OPN) and vimentin) in pulmonary artery was restored by icotinib treatment. In vitro, icotinib suppressed EGF-induced PASMCs proliferation and migration. Meanwhile, icotinib inhibited EGF-induced downregulation of α-smooth muscle actin and SM22α and upregulation of OPN and Collagen I in PASMCs, suggesting that icotinib could inhibit EGF-induced phenotypic switching of PASMCs. Mechanistically, these effects of icotinib were associated with the inhibition of EGFR-Akt/ERK signaling pathway. CONCLUSIONS: Icotinib can attenuate MCT-induced pulmonary vascular remodeling and improve PH. This effect of icotinib might be attributed to preventing PASMC dysfunction by inhibiting EGFR-Akt/ERK signaling pathway.
Asunto(s)
Éteres Corona/farmacología , Receptores ErbB/antagonistas & inhibidores , Hipertensión Pulmonar/fisiopatología , Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Arteria Pulmonar/efectos de los fármacos , Quinazolinas/farmacología , Animales , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Factor de Crecimiento Epidérmico/farmacología , Hipertensión Pulmonar/inducido químicamente , Técnicas In Vitro , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas de Microfilamentos/efectos de los fármacos , Proteínas de Microfilamentos/metabolismo , Monocrotalina/toxicidad , Proteínas Musculares/efectos de los fármacos , Proteínas Musculares/metabolismo , Músculo Liso Vascular/fisiopatología , Osteopontina/efectos de los fármacos , Osteopontina/metabolismo , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Arteria Pulmonar/fisiopatología , Ratas , Transducción de Señal , Remodelación Vascular/efectos de los fármacos , Función Ventricular Derecha/efectos de los fármacos , Presión Ventricular/efectos de los fármacos , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
Mesenchymal stem cell (MSC)-based therapies have been used in skin regeneration due to their ability to differentiate into many cells, promote cytokine secretion and participate in collagen deposition. In this study, we concluded that a CuS@BSA nanoparticles exhibited similar potential in inducing MSCs differentiation to fibroblasts as Cu ions for wound healing. Methods: First, we verified the photothermal efficiency of CuS@BSA in vivo and vitro and had no cytotoxicity for MSCs when the temperature was controlled at 42 °C by adjusting the power of irradiation at 980 nm. And then we detected the expression of vimentin in MSCs, which further directed the MSCs to fibroblasts through Western blotting and Immunofluorescence when treated with CuS@BSA or pre-heat at 42 °C. In addition, we implanted MSCs into the Matrigel or electrospun PLA nanofiber membrane in vitro to evaluating the effect of heating or CuS@BSA on the morphological change of MSCs by SEM. Finally, we evaluated improving skin regeneration by the combination of preheated-MSCs and CuS@BSA nanoparticles that were encapsulated in Matrigel. Results: The CuS@BSA nanoparticles have good photothermal conversion efficiency. Not only CuS nanoparticles itself or after irradiation at 980 nm stimulated the expressioin of vimentin in MSCs. Besides, the CuS@BSA can promote cell proliferation as Cu ion through the expression of ERK. The combination of the CuS@BSA nanoparticles and thermal treatment synergistically improved the closure of an injured wound in an injured wound model. Conclusions: MSCs combined with CuS@BSA are a promising wound dressing for the reconstruction of full-thickness skin injuries.
Asunto(s)
Cobre/farmacología , Fibroblastos/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de los fármacos , Regeneración/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Cobre/administración & dosificación , Fibroblastos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Nanopartículas/administración & dosificación , Fototerapia/métodos , Ratas , Ratas Sprague-Dawley , Regeneración/fisiología , Piel/efectos de los fármacos , Piel/lesiones , Vimentina/biosíntesis , Vimentina/efectos de los fármacosRESUMEN
BACKGROUND: Epithelial-mesenchymal transformation (EMT) is a central mechanism for the occurrence and development of pulmonary fibrosis. Therefore, to identify the key target molecules regulating the EMT process is considered as an important direction for the prevention and treatment of pulmonary fibrosis. Transglutaminase 2 (TG2) has been recently found to play an important role in the regulation of inflammation and the generation of extracellular matrix. Here, our study focuses on the roles of TG2 in pulmonary fibrosis and EMT. METHODS: at first, the expression of TG2 and the EMT-related markers like E-cadherin, Vimentin, and α-SMA were detected with Western Blotting, immunohistochemistry and other methods in the mice with pulmonary fibrosis induced by bleomycin. Further, MLE 12 cells were used to study the effects on EMT of the inhibition of TG2 in vitro. Finally, GK921, an inhibitor against TG2, was used to show its function in both prevention and treatment of pulmonary fibrosis induced by bleomycin in mice. RESULTS: bleomycin succeeded to induce pulmonary fibrosis in mice, with increased TG2 expression, EMT and Akt activation. Knock-down of TG2 by siRNA technique in MLE 12 cell (a mouse alveolar epithelial cell line) and GK921 (an inhibitor of TG2) all inhibited the EMT process, however SC79, an activator of Akt rescued above inhibition. Finally, GK921 alleviated pulmonary fibrosis in mice induced by bleomycin. CONCLUSION: Blocking TG2 reduces bleomycin-induced pulmonary fibrosis in mice via inhibiting EMT.
Asunto(s)
Transición Epitelial-Mesenquimal/fisiología , Proteínas de Unión al GTP/genética , Fibrosis Pulmonar/genética , Transglutaminasas/genética , Acetatos/farmacología , Actinas/efectos de los fármacos , Actinas/metabolismo , Animales , Antibióticos Antineoplásicos/toxicidad , Benzopiranos/farmacología , Bleomicina/toxicidad , Cadherinas/efectos de los fármacos , Cadherinas/metabolismo , Línea Celular , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Transición Epitelial-Mesenquimal/efectos de los fármacos , Proteínas de Unión al GTP/antagonistas & inhibidores , Proteínas de Unión al GTP/metabolismo , Técnicas de Silenciamiento del Gen , Técnicas In Vitro , Ratones , Proteína Glutamina Gamma Glutamiltransferasa 2 , Fibrosis Pulmonar/metabolismo , Pirazinas/farmacología , Mucosa Respiratoria/citología , Transglutaminasas/antagonistas & inhibidores , Transglutaminasas/metabolismo , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
Rikkunshito, a traditional Japanese herbal medicine, improves appetite via activation of gastrointestinal hormone ghrelin pathway. The function of ghrelin is mediated by growth hormone secretagogue receptor (GHSR1a), and ghrelin has been known to possess diverse physiological functions including growth suppression of some cancer cells. Considering that increased ghrelin signaling by Rikkunshito could enhance sirtuin1 (SIRT1) activity in nervous system, we aimed to investigate the effect of Rikkunshito in ovarian cancer cells. Ovarian cancer cell lines were treated with Rikkunshito, and cellular viability, gene expressions and epithelial-mesenchymal transition (EMT) status were investigated. To investigate the involvement of SIRT1 by Rikkunshito in SKOV3 cancer cells, endogenous expression of SIRT1 was depleted using small interfering RNA (siRNA). Treatment with Rikkunshito elevated ghrelin, GHSR1a and SIRT1, while cellular viability was decreased. The treatment of Rikkunshito also inhibited cellular migration and invasion status in a dose-dependent manner, and these effects were translated to the enhanced EMT status, although the role of SIRT1 was not determined. Our study revealed a novel function of Rikkunshito in enhancing EMT status of ovarian cancer cells. Therefore, we would like to propose that Rikkunshito may be used as a novel adjunctive therapy in chemotherapy of ovarian cancer because platinum-based chemotherapy frequently used for the treatment of ovarian cancer inevitably impairs appetite.
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Medicamentos Herbarios Chinos/farmacología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Neoplasias Ováricas/metabolismo , Receptores de Ghrelina/efectos de los fármacos , Sirtuina 1/efectos de los fármacos , Antígenos CD/efectos de los fármacos , Antígenos CD/metabolismo , Cadherinas/efectos de los fármacos , Cadherinas/metabolismo , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Supervivencia Celular/efectos de los fármacos , Transición Epitelial-Mesenquimal/genética , Femenino , Expresión Génica/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Ghrelina/efectos de los fármacos , Ghrelina/metabolismo , Humanos , Neoplasias Ováricas/genética , Receptores de Ghrelina/genética , Receptores de Ghrelina/metabolismo , Sirtuina 1/genética , Sirtuina 1/metabolismo , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
BACKGROUND Colorectal cancer is one of the leading causes of death in China, and the development of effective drugs is urgently needed. Here, we report on Paeoniflorin (PF), a product isolated from the roots of the peony plant, as a possible candidate because of its anti-tumor effects on epithelial-to-mesenchymal transition (EMT) of PF in human colorectal cancer (CRC). MATERIAL AND METHODS Cell proliferation, wound healing, and Transwell assays were used to analyze the effects of PF on in vitro cell migration and invasion of HCT116 and SW480, 2 colorectal cancer cell lines. The tumor xenograft model was used to verify the anti-metastasis effects of PF in vivo. The RNA and protein levels of epithelia-cadherin (E-cadherin), Vimentin, and histone deacetylase2 (HDAC2) were measured by qPCR and Western blot analysis to explore the mechanism involved. RESULTS Our results showed that PF inhibited colorectal cancer cell migration and invasion and suppressed the metastatic potential of the cancer cells in vivo. Moreover, PF significantly decreased the expression of HDAC2 and Vimentin, while increasing the expression of E-cadherin. CONCLUSIONS These results suggest that PF inhibits colorectal cancer cell migration and invasion ability and reverses the EMT process, through inhibiting the expression of HDAC2, and then affects the expression level of E-cadherin and Vimentin at the cell level. Our results were also verified in the tumor xenograft model. This indicates that PF may be a candidate for colorectal cancer treatment.
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Neoplasias Colorrectales/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Glucósidos/farmacología , Monoterpenos/farmacología , Animales , Cadherinas/efectos de los fármacos , Línea Celular Tumoral/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , China , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Histona Desacetilasa 2/efectos de los fármacos , Humanos , Medicina Tradicional China , Ratones , Ratones Desnudos , Transducción de Señal/efectos de los fármacos , Vimentina/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Improved biocompatibility of peritoneal dialysis solution (PDS) is crucial for peritoneal membrane preservation, thereby ensuring long-term peritoneal dialysis (PD) and preventing encapsulating peritoneal sclerosis. We previously reported the protective effect of molecular hydrogen (H2) on mesothelial cells from PDS in nonuremic rats. SUMMARY: In the present study, we examined the effect of H2-containing PDS (commercially available neutral pH type) regarding the protection of peritoneal tissue in experimental chronic kidney disease rats. Furthermore, we conducted a 2-week clinical trial in which H2-containing PDS was used in place of standard PDS and its feasibility was examined. In the experimental study, test solutions were injected through the subcutaneous port into the abdomen for 3 weeks. Histological study revealed a significant increase in the number of mesothelial cells and a significant decrease in peritoneal thickness in the H2-PD group as compared to the control and PD groups. Also, results of immunostaining analysis revealed increased vimentin and apoptotic cells in the membrane of the PD group, indicating that H2 may play a role in ameliorating PDS-induced peritoneal injury and preserving peritoneal integrity. In the clinical trial with 6 prevalent PD patients, all subjects completed the study with no adverse effects. Moreover, there were substantial changes in surrogate markers, such as increased CA125 and mesothelin, in the effluent in selected cases, suggesting enhanced mesothelial regeneration by H2. Key Message: H2-enriched PDS is a candidate novel PDS with improved biocompatibility. Further, our results support the significance of H2-PD clinical trials in the future.
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Soluciones para Diálisis/química , Hidrógeno/farmacología , Diálisis Peritoneal/métodos , Animales , Apoptosis , Células Epiteliales/citología , Epitelio/efectos de los fármacos , Epitelio/fisiología , Humanos , Hidrógeno/uso terapéutico , Ratas , Regeneración/efectos de los fármacos , Investigación Biomédica Traslacional , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
Diabetes mellitus (DM) affects many organs including kidney. Tyrosine kinase can cause hypoglycemia and sunitinib is an inhibitor of tyrosine kinase. We investigated the possible effects of sunitinib on the kidney of streptozotocin (STZ) induced type 1 diabetic mice. We used 28 CD 1 type male mice divided into four groups of seven. Type 1 diabetes was induced by injection of STZ. Group 1 was the untreated control. Group 2 comprised non-diabetic mice + sunitinib. Both groups 1 and 2 exhibited normal blood glucose levels. Group 3 comprised STZ treated diabetic mice + saline. Group 4 were diabetic mice + sunitinib treatment. Kidneys were removed after 8 weeks. The immunoreactivities of vimentin, E-cadherin and S100 were assessed. Immunostaining of vimentin, E-cadherin and S100 was located in both the glomeruli and tubules of the kidney. We found that the number of vimentin and E-cadherin positive glomeruli and tubules were increased after sunitinib treatment compared to saline treated diabetic mice. The number of vimentin labeled tubules was decreased in the sunitinib treated group compared to diabetic + saline groups. Differences in the number of S100 positive tubules and glomeruli between groups 3 and 4 were not statistically significant. The effect of sunitinib on experimental diabetic mice appears to be related to levels of vimentin, E-cadherin and S100 in the glomeruli and tubules of the kidney, and sunitinib may protect against renal damage from DM.
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Cadherinas/efectos de los fármacos , Nefropatías Diabéticas/metabolismo , Proteínas S100/efectos de los fármacos , Sunitinib/farmacología , Vimentina/efectos de los fármacos , Animales , Cadherinas/metabolismo , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Tipo 1/metabolismo , Nefropatías Diabéticas/tratamiento farmacológico , Riñón/metabolismo , Glomérulos Renales/metabolismo , Masculino , Ratones , Proteínas S100/metabolismo , Estreptozocina/farmacología , Vimentina/metabolismoRESUMEN
Sertoli cells (SCs) provide physical and nutritional support for spermatogenesis. Dibutyl phthalate (DBP) is a plasticizer that has male reproductive toxicity. The collapse of vimentin in DBP-exposed SCs is thought to induce the sloughing of spermatocytes from seminiferous tubules. In this study, we explored methods to rescue vimentin from collapse in DBP-exposed SCs. DBP not only induced the hyperphosphorylation of vimentin but also triggered endoplasmic reticulum (ER) stress and apoptosis in SCs. Treatment with BAPTA-AM, an antagonist of Ca2+, significantly decreased the level of phosphorylated vimentin, while LY294002, an inhibitor of Akt1, did not. ER stress and apoptosis remained at high levels, and the distribution of vimentin was not improved. ZnSO4 treatment did not decrease the level of phosphorylated vimentin. However, after treatment, ER stress and apoptosis were obviously inhibited, and the distribution of vimentin was reconverted. These results indicated that ZnSO4 could alleviate the collapse of vimentin by attenuating ER stress and apoptosis. This study suggested that an appropriate zinc supply might be a choice to alleviate DBP-induced adverse reproductive effects.
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Apoptosis/efectos de los fármacos , Dibutil Ftalato/toxicidad , Estrés del Retículo Endoplásmico/efectos de los fármacos , Plastificantes/toxicidad , Células de Sertoli/efectos de los fármacos , Vimentina/metabolismo , Sulfato de Zinc/farmacología , Animales , Proteínas Reguladoras de la Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Quelantes/farmacología , Niño , Cromonas/farmacología , Ácido Egtácico/análogos & derivados , Ácido Egtácico/farmacología , Proteínas de Choque Térmico/metabolismo , Humanos , Masculino , Morfolinas/farmacología , Fosforilación , Cultivo Primario de Células , Ratas , Ratas Sprague-Dawley , Vimentina/efectos de los fármacosRESUMEN
OBJECTIVE: To investigate the safety, tolerability, pharmacokinetics, and pharmacodynamics of ABT-981, a human dual variable domain immunoglobulin simultaneously targeting interleukin (IL)-1α and IL-1ß, in patients with knee osteoarthritis (OA). METHOD: This was a randomized, double-blind, placebo-controlled, single-center study of multiple subcutaneous (SC) injections of ABT-981 in patients with mild-to-moderate OA of the knee (NCT01668511). Three cohorts received ABT-981 (0.3, 1, or 3 mg/kg) or placebo every other week for a total of four SC injections, and one cohort received ABT-981 (3 mg/kg) or placebo every 4 weeks for a total of three SC injections. Assessment of safety and tolerability were the primary objectives. A panel of serum and urine biomarkers of inflammation and joint degradation were evaluated. RESULTS: A total of 36 patients were randomized (ABT-981, n = 28; placebo, n = 8); 31 (86%) completed the study. Adverse event (AE) rates were comparable between ABT-981 and placebo (54% vs 63%). The most common AE reported with ABT-981 vs placebo was injection site erythema (14% vs 0%). ABT-981 significantly reduced absolute neutrophil count and serum concentrations of IL-1α/IL-1ß, high-sensitivity C-reactive protein, and matrix metalloproteinase (MMP)-derived type 1 collagen. Serum concentrations of MMP-derived type 3 collagen and MMP-degraded C-reactive protein demonstrated decreasing trends with ABT-981. Antidrug antibodies were found in 37% of patients but were not associated with the incidence or severity of AEs. CONCLUSION: ABT-981 was generally well tolerated in patients with knee OA and engaged relevant tissue targets, eliciting an anti-inflammatory response. Consequently, ABT-981 may provide clinical benefit to patients with inflammation-driven OA.
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Inmunoglobulinas/uso terapéutico , Interleucina-1alfa/antagonistas & inhibidores , Interleucina-1beta/antagonistas & inhibidores , Osteoartritis de la Rodilla/tratamiento farmacológico , Anciano , Agrecanos/efectos de los fármacos , Agrecanos/metabolismo , Proteína C-Reactiva/efectos de los fármacos , Proteína C-Reactiva/metabolismo , Proteína de la Matriz Oligomérica del Cartílago/efectos de los fármacos , Proteína de la Matriz Oligomérica del Cartílago/metabolismo , Citrulinación , Colágeno Tipo I/efectos de los fármacos , Colágeno Tipo I/metabolismo , Colágeno Tipo II/efectos de los fármacos , Colágeno Tipo II/metabolismo , Colágeno Tipo III/efectos de los fármacos , Colágeno Tipo III/metabolismo , Eritema , Femenino , Humanos , Inmunoglobulinas/farmacología , Reacción en el Punto de Inyección , Inyecciones Subcutáneas , Interleucina-1beta/efectos de los fármacos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Neutrófilos/citología , Osteoartritis de la Rodilla/metabolismo , Péptidos/efectos de los fármacos , Péptidos/metabolismo , Índice de Severidad de la Enfermedad , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/metabolismo , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
OBJECTIVE: To compare the proliferative and periodontal specific differentiation abilities of induced pluripotent stem cells (iPSCs) at different passages, and to investigate whether long term culturing would have a negative influence on their proliferation and specific differentiation capacity, thus providing a theoretical basis for further in-depth research on periodontal regeneration and the possible clinical applications of iPSCs. METHODS: IPSCs derived from human gingival fibroblasts at passages 5, 10, 15 and 20 were recovered and cultured in vitro. Their morphology and proliferation rates were observed respectively. We further induced the iPSCs at different passages toward periodontal tissue under the treatment of growth/differentiation factor-5 (GDF-5) for 14 days through the EB routine, then compared the periodontal differentiation propensities between the different passages of iPSCs by detecting their calcified nodules formation by Alizarin red staining and assaying their relative periodontal tissue related marker expressions by qRT-PCR and immunofluorescence staining, including bone related markers: osteocalcin (OCN), bone sialoprotein (BSP); periodontal ligament related markers: periostin, vimentin; and cementum related markers: cementum attachment protein (CAP), cementum protein 1 (CEMP1). The untreated spontaneous differentiation groups were set as negative controls respectively. RESULTS: iPSCs at different passages all showed a high proliferative capacity when cultured in vitro and turned into a spindle-like shape similar to fibroblasts upon periodontal specific differentiation. All iPSCs formed typical calcified nodules upon GDF-5 induction by Alizarin red staining in comparison to their untreated controls. The relative calcium deposition at all passages had been significantly upgraded under the treatment of GDF-5 (P5: t=2.125, P=0.003; P10: t=2.246, P=0.021; P15: t=3.754, P=0.004; P20: t=3.933, P=0.002), but no significant difference in their calcium deposition were detected within passages 5, 10, 15 and 20 (periodontal differentiation: F=2.365, P=0.109; spontaneously differentiation: F=2.901, P=0.067). Periodontal tissue related marker expressions of iPSCs at all passages had also been significantly upgraded under the treatment of GDF-5 (P<0.05), but still, no significant difference in their expression levels of periodontal tissue related proteins were detected within passages (BSP: F=0.926 7, P=0.450; vimentin: F=0.917 1, P=0.455; CEMP1: F=2.129, P=0.1367). CONCLUSION: Our results preliminarily confirmed that long term culturing won't influence the proliferation capacity and periodontal specific differentiation propensity of iPSCs, as they can still proliferate and differentiate toward periodontal cells with high efficiency upon growth factor induction after continuous passaging. Therefore, iPSCs could be recognized as a promising cell source for future possible application in periodontal tissue regeneration.
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Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Células Cultivadas/fisiología , Células Madre Pluripotentes Inducidas/fisiología , Calcificación Fisiológica/efectos de los fármacos , Calcificación Fisiológica/genética , Moléculas de Adhesión Celular/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Fibroblastos/efectos de los fármacos , Encía , Factor 5 de Diferenciación de Crecimiento/farmacología , Humanos , Sialoproteína de Unión a Integrina/efectos de los fármacos , Sialoproteína de Unión a Integrina/metabolismo , Osteocalcina/efectos de los fármacos , Osteocalcina/metabolismo , Periodoncio/efectos de los fármacos , Periodoncio/crecimiento & desarrollo , Proteínas Tirosina Fosfatasas/efectos de los fármacos , Proteínas Tirosina Fosfatasas/metabolismo , Proteínas/efectos de los fármacos , Proteínas/metabolismo , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
Objective: To investigate the effect of a novel EZH2 inhibitor GSK126 on cell growth, apoptosis and migration of prostate cancer cells. Methods: Prostate cancer PC-3 and DU145 cells were treated with GSK126 at different doses. Cell growth was detected by sulforhodamine assay. Cell apoptosis was assayed by Annexin V-/PI kit. Transwell chamber and wound healing assays were conducted to detect cell migration. The mRNA level was detected by quantitative PCR, and protein expression was detected by Western blot analysis. Results: GSK126 showed significant effect on cell growth and apoptosis when the dose was higher than 50 µmol/L. Wound healing assay revealed that scratch space in PC-3 cells was significantly increased in a dose-dependent manner in GSK126-treated groups[(247.2±24.4),(347.2±19.2) and (410.5±18.1) µm in low, medium and high dose (5.0, 20.0, 50.0 µmol/L), respectively] as compared with the control group[(171.3±17.8) µm](all P<0.05). Transwell assay showed that migrated PC-3 cells in control group was 322.0±17.9,while those in GSK126-treated groups were 198.3±15.4 (low),82.7±6.2 (medium) and 30.2±4.1 (high), and the differences between the control group and GSK126-treated groups were significant(all P<0.05). In addition, GSK126 up-regulated E-cadherin mRNA expression and down-regulated N-cadherin and Vimentin mRNA expression, whereas had no significant effect on Snail, Fibronectin and VEGF-A mRNA expression. The protein expression of E-cadherin was elevated but VEGF-A protein did not change in GSK126-treated groups. Similar results were exhibited in DU145 cell. Conclusion: GSK126 can significantly inhibit cell migration and invasion in prostate cancer PC-3 and DU145 cells, which may be resulted from its effect on epithelial-mesenchymal transition. GSK126 may be used as a potential anti-prostate cancer dug in clinic.
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Indoles/farmacología , Neoplasias de la Próstata/fisiopatología , Piridonas/farmacología , Antígenos CD , Apoptosis/efectos de los fármacos , Cadherinas/análisis , Cadherinas/efectos de los fármacos , Cadherinas/metabolismo , Línea Celular Tumoral/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales/métodos , Proteína Potenciadora del Homólogo Zeste 2/análisis , Proteína Potenciadora del Homólogo Zeste 2/efectos de los fármacos , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Fibronectinas/análisis , Fibronectinas/efectos de los fármacos , Fibronectinas/metabolismo , Humanos , Masculino , Neoplasias de la Próstata/química , Neoplasias de la Próstata/genética , ARN Mensajero , Regulación hacia Arriba/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/análisis , Factor A de Crecimiento Endotelial Vascular/efectos de los fármacos , Vimentina/análisis , Vimentina/efectos de los fármacos , Vimentina/metabolismoRESUMEN
The sonic hedgehog (Shh) pathway is highly activated in thyroid neoplasms and promotes thyroid cancer stem-like cell phenotype, but whether the Shh pathway regulates thyroid tumor cell motility and invasiveness remains unknown. Here, we report that the motility and invasiveness of two anaplastic thyroid tumor cell lines, KAT-18 and SW1736, were inhibited by two inhibitors of the Shh pathway (cyclopamine and GANT61). Consistently, the cell motility and invasiveness was decreased by Shh and Gli1 knockdown, and was increased by Gli1 overexpression in KAT-18 cells. Mechanistic studies revealed that Akt and c-Met phosphorylation was decreased by a Gli1 inhibitor and by Shh and Gli1 knockdown, but was increased by Gli1 overexpression. LY294002, a PI-3 kinase inhibitor, and a c-Met inhibitor inhibited the motility and invasiveness of Gli1-transfected KAT-18 cells more effectively than the vector-transfected cells. Knockdown of Snail, a transcription factor regulated by the Shh pathway, led to decreased cell motility and invasiveness in KAT-18 and SW1736 cells. However, key epithelial-to-mesenchymal transition (EMT) markers including E-cadherin and vimentin as well as Slug were not affected by cyclopamine and GANT61 in either SW1736 or WRO82, a well differentiated follicular thyroid carcinoma cell line. Our data suggest that the Shh pathway-stimulated thyroid tumor cell motility and invasiveness is largely mediated by AKT and c-Met activation with little involvement of EMT.