RESUMO
During intercellular communication, cells release extracellular vesicles such as exosomes, which contain proteins, ncRNAs and mRNAs that can influence proliferation and/or trigger apoptosis in recipient cells, and have been proposed to play an essential role in promoting invasion of tumor cells and in the preparation of metastatic niches. Our group proposed the antisense non-coding mitochondrial RNA (ASncmtRNA) as a new target for cancer therapy. ASncmtRNA knockdown using an antisense oligonucleotide (ASO-1537S) causes massive death of tumor cells but not normal cells and strongly reduces metastasis in mice. In this work, we report that exosomes derived from ASO-1537S-treated MDA-MB-231 breast cancer cells (Exo-1537S) inhibits tumorigenesis of recipient cells, in contrast to exosomes derived from control-ASO-treated cells (Exo-C) which, in contrast, enhance these properties. Furthermore, an in vivo murine peritoneal carcinomatosis model showed that Exo-1537S injection reduced tumorigenicity compared to controls. Proteomic analysis revealed the presence of Lactadherin and VE-Cadherin in exosomes derived from untreated cells (Exo-WT) and Exo-C but not in Exo-1537S, and the latter displayed enrichment of proteasomal subunits. These results suggest a role for these proteins in modulation of tumorigenic properties of exosome-recipient cells. Our results shed light on the mechanisms through which ASncmtRNA knockdown affects the preparation of breast cancer metastatic niches in a peritoneal carcinomatosis model.
Assuntos
Exossomos/metabolismo , Mitocôndrias/genética , RNA não Traduzido/metabolismo , Animais , Antígenos CD/metabolismo , Antígenos de Superfície/metabolismo , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Caderinas/metabolismo , Carcinogênese/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Humanos , Camundongos , Proteínas do Leite/metabolismo , Oligorribonucleotídeos Antissenso/metabolismo , Oligorribonucleotídeos Antissenso/farmacologia , RNA não Traduzido/antagonistas & inibidores , RNA não Traduzido/genética , Transplante HeterólogoRESUMO
Breast cancer is one of the most frequently diagnosed cancers and the leading cause of cancer-related deaths in women worldwide, whereby mortality is largely attributable to the development of distant metastasis. Caveolin-1 (CAV1) is a multifunctional membrane protein that is typically upregulated in the final stages of cancer and promotes migration and invasion of tumor cells. Elevated levels of CAV1 have been detected in extracellular vesicles (EVs) from advanced cancer patients. EVs are lipid enclosed vesicular structures that contain bioactive proteins, DNA and RNAs, which can be transferred to other cells and promote metastasis. Therefore, we hypothesized that CAV1 containing EVs released from breast cancer cells may enhance migration and invasion of recipient cells. EVs were purified from conditioned media of MDA-MB-231 wild-type (WT), MDA-MB-231 (shCAV1; possessing the plasmid pLKO.1 encoding a 'small hairpin' directed against CAV1) and MDA-MB-231 (shC) short hairpin control cells. Nanoparticle tracking analysis revealed an average particle size of 40-350 nm for all preparations. As anticipated, CAV1 was detected in MDA-MB-231 WT and shC EVs, but not in MDA-MB-231 (shCAV1) EVs. Mass spectrometry analysis revealed the presence of specific cell adhesion-related proteins, such as Cyr61, tenascin (TNC) and S100A9 only in WT and shC, but not in shCAV1 EVs. Importantly, EVs containing CAV1 promoted migration and invasion of cells lacking CAV1. We conclude that the presence of CAV1 in EVs from metastatic breast cancer cells is associated with enhanced migration and invasiveness of recipient cells in vitro, suggesting that intercellular communication promoted by EVs containing CAV1 will likely favor metastasis in vivo.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Caveolina 1/genética , Adesão Celular/efeitos dos fármacos , Neoplasias da Mama/química , Neoplasias da Mama/patologia , Caveolina 1/química , Comunicação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Vesículas Extracelulares/química , Feminino , Humanos , Células MCF-7 , Metástase NeoplásicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Ibervillea sonorae (S. Watson) Greene (Cucurbitaceae), a plant used for the empirical treatment of type 2 diabetes in México, exerts antidiabetic effects on animal models but its mechanism of action remains unknown. The aim of this study is to investigate the antidiabetic mechanism of an Ibervillea sonorae aqueous extract (ISE). MATERIALS AND METHODS: Non-toxic ISE concentrations were assayed on the glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes, both in the absence or the presence of insulin signaling pathway inhibitors, and on murine and human adipogenesis. Chemical composition of ISE was examined by spectrophotometric and HPLC techniques. RESULTS: ISE stimulated the 2-NBDGlucose uptake by mature adipocytes in a concentration-dependent manner. ISE 50 µg/ml induced the 2-NBDG uptake in insulin-sensitive 3T3-F442A, 3T3-L1 and human adipocytes by 100%, 63% and 33%, compared to insulin control. Inhibitors for the insulin receptor, PI3K, AKT and GLUT4 blocked the 2-NBDG uptake in murine cells, but human adipocytes were insensitive to the PI3K inhibitor Wortmannin. ISE 50 µg/ml also stimulated the 2-NBDG uptake in insulin-resistant adipocytes by 117% (3T3-F442A), 83% (3T3-L1) and 48% (human). ISE induced 3T3-F442A adipogenesis but lacked proadipogenic effects on 3T3-L1 and human preadipocytes. Chemical analyses showed the presence of phenolics in ISE, mainly an appreciable concentration of gallic acid. CONCLUSION: Ibervillea sonorae exerts its antidiabetic properties by means of hydrosoluble compounds stimulating the glucose uptake in human preadipocytes by a PI3K-independent pathway and without proadipogenic effects.
Assuntos
Adipócitos/efeitos dos fármacos , Cucurbitaceae/química , Hipoglicemiantes/farmacologia , Extratos Vegetais/farmacologia , Células 3T3 , Adipócitos/metabolismo , Adipogenia/efeitos dos fármacos , Animais , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Glucose/metabolismo , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/isolamento & purificação , Insulina/metabolismo , Resistência à Insulina , Medicina Tradicional , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Transdução de Sinais/efeitos dos fármacos , EspectrofotometriaRESUMO
Some Magnolia (Magnoliaceae) species are used for the empirical treatment of diabetes mellitus, but the antidiabetic properties of Magnolia dealbata have not yet been experimentally validated. Here we report that an ethanolic extract of Magnolia dealbata seeds (MDE) and its active principles honokiol (HK) and magnolol (MG) induced the concentration-dependent 2-NBDG uptake in murine 3T3-F442A and human subcutaneous adipocytes. In insulin-sensitive adipocytes, MDE 50 µg/ml induced the 2-NBDG uptake by 30% respect to insulin, while HK and MG, 30 µM each, did it by 50% (murine) and 40% (human). The simultaneous application of HK and MG stimulated 2-NBDG uptake by 70% in hormone-sensitive cells, on which Magnolia preparations exerted synergic effects with insulin. In insulin-resistant adipocytes, MDE, HK and MG induced 2-NBDG uptake by 57%, 80% and 96% respect to Rosiglitazone (RGZ), whereas HK and MG simultaneously applied stimulated 2-NBDG uptake more efficiently than RGZ (120%) in both murine and human adipocytes. Inhibitors of the insulin-signaling pathway abolished the glucose uptake induced by Magnolia dealbata preparations, suggesting that their antidiabetic effects are mediated by this signaling pathway. In addition, MDE, HK and MG exerted only mild to moderate proadipogenic effects on 3T3-F442A and human preadipocytes, although the combined application of HK and MG markedly increased the lipid accumulation in both cell types. In summary, Magnolia dealbata and its active principles HK and MG stimulate glucose uptake in insulin-sensitive and insulin-resistant murine and human adipocytes using the insulin signaling pathway.
Assuntos
Adipócitos/efeitos dos fármacos , Compostos de Bifenilo/farmacologia , Glucose/metabolismo , Lignanas/farmacologia , Magnolia/química , Transdução de Sinais , Células 3T3 , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/metabolismo , Adipogenia , Animais , Sobrevivência Celular/efeitos dos fármacos , Desoxiglucose/análogos & derivados , Desoxiglucose/metabolismo , Sinergismo Farmacológico , Humanos , Hipoglicemiantes/farmacologia , Insulina/metabolismo , Metabolismo dos Lipídeos , Camundongos , Extratos Vegetais/farmacologiaRESUMO
The antimicrobial effects of the Mexican medicinal plants Guazuma ulmifolia, Justicia spicigera, Opuntia joconostle, O. leucotricha, Parkinsonia aculeata, Phoradendron longifolium, P. serotinum, Psittacanthus calyculatus, Tecoma stans and Teucrium cubense were tested against several human multi-drug resistant pathogens, including three Gram (+) and five Gram (-) bacterial species and three fungal species using the disk-diffusion assay. The cytotoxicity of plant extracts on human cancer cell lines and human normal non-cancerous cells was also evaluated using the MTT assay. Phoradendron longifolium, Teucrium cubense, Opuntia joconostle, Tecoma stans and Guazuma ulmifolia showed potent antimicrobial effects against at least one multidrug-resistant microorganism (inhibition zone > 15 mm). Only Justicia spicigera and Phoradendron serotinum extracts exerted active cytotoxic effects on human breast cancer cells (IC50 < or = 30 microg/mL). The results showed that Guazuma ulmifolia produced potent antimicrobial effects against Candida albicans and Acinetobacter lwoffii, whereas Justicia spicigera and Phoradendron serotinum exerted the highest toxic effects on MCF-7 and HeLa, respectively, which are human cancer cell lines. These three plant species may be important sources of antimicrobial and cytotoxic agents.
Assuntos
Anti-Infecciosos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Extratos Vegetais/farmacologia , Plantas Medicinais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , MéxicoRESUMO
AIM OF THE STUDY: Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae) and Teucrium cubense Jacq (Lamiaceae) are plants extensively used for the empirical treatment of diabetes mellitus, but their antidiabetic mechanisms remain to be clarified. In this study, the effect of aqueous extracts of Tecoma stans (TSE) and Teucrium cubense (TCE) on the glucose uptake in adipose cells was evaluated. MATERIALS AND METHODS: Non-toxic concentrations of TSE and TCE were assayed on the adipogenesis and 2-NBDglucose uptake in insulin-sensitive and insulin-resistant murine 3T3-F442A and human subcutaneous adipocytes. RESULTS: Both extracts stimulated 2-NBDG uptake by insulin-sensitive and insulin-resistant adipocytes in a concentration-dependent manner. In insulin-sensitive cells, TSE 70 microg/ml stimulated 2-NBDG uptake by 193% (murine) and by 115% (human), whereas the same concentration of TCE induced the 2-NBDG uptake by 112% (murine) and 54% (human). In insulin-resistant adipocytes, TSE induced the 2-NBDG uptake by 94% (murine) and 70% (human), compared with the incorporation shown by insulin-sensitive adipocytes stimulated by the hormone, whereas TCE induced the incorporation of 2-NBDG by 69% (murine) and 31% (human). On the other hand, TSE and TCE exerted only minimal or null proadipogenic effects on murine and human preadipocytes. CONCLUSION: Tecoma stans and Teucrium cubense exert their antidiabetic effects stimulating glucose uptake in both insulin-sensitive and insulin-resistant murine and human adipocytes without significant proadipogenic or antiadipogenic side effects.