RESUMO
Zoledronic acid (ZA), a third-generation bisphosphonate, has been extensively used to treat osteoporosis and cancer bone metastasis and demonstrated to suppress proliferation of varied cancer cells and selectively kill tumor-associated microphages (TAMs). However, the clinical applications of ZA in extraskeletal tumor treatment are largely restricted due to its rapid renal clearance and binding to bones. In this study, to promote intracellular delivery of ZA for amplified antitumor efficacy, tumor acidity-responsive polymeric nanoparticles with high ZA payload (ca. 12.3 wt%) and low premature ZA leakage were designed. As a pivotal material for surface coating, the acidity-sensitive and amphiphilic methoxy poly(ethylene glycol) (mPEG)-benzoic imine-octadecane (C18) (mPEG-b-C18) was synthesized by conjugation of mPEG-CHO with 1-octadecylamine upon Schiff base reaction. Through tailor-made co-assembly of the hydrophobic poly(lactic-co-glycolic acid) (PLGA), amphiphilic tocopheryl polyethylene glycol succinate (TPGS) and mPEG-b-C18 to encapsulate ionic complexes composed of ZA molecules and branched poly(ethylenimine) (PEI) segments, the attained therapeutic polymeric nanoparticles, characterized to have a hydrophobic PLGA/ZA/PEI-constituted core covered with mPEG-b-C18 and TPGS, were able to not only detach mPEG shielding upon acidity-triggered hydrolysis of benzoic imine bonds but also expose surface positive charges of protonated PEI segments. The in vitro cellular uptake and cytotoxicity studies demonstrated that the internalization of acidity-sensitive ZA-encapsulated nanoparticles by TRAMP-C1 mouse prostate cancer cells and murine macrophages RAW 264.7 was considerably promoted upon acidity-elicited PEG detachment and surface charge conversion, thus remarkably boosting intracellular ZA delivery and anticancer potency. Compared to PEG non-detachable ZA-loaded nanoparticles with poor tumor deposition and antitumor effect, the PEG-detachable ZA-carrying nanoparticles markedly accumulated in TRAMP-C1 solid tumors in vivo and inhibited tumor growth, thereby increasing the survival rate of the treated mice. The collective data suggest the great promise of tumor acidity-sensitive ZA-carrying hybrid nanoparticles in the treatment of extraskeletal solid tumors.
Assuntos
Nanopartículas , Neoplasias , Polietilenoglicóis/química , Animais , Iminas , Masculino , Camundongos , Nanopartículas/química , Neoplasias/tratamento farmacológico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/uso terapêutico , Polímeros/química , Ácido Zoledrônico/farmacologia , Ácido Zoledrônico/uso terapêuticoRESUMO
To achieve effective intracellular anticancer drug release for boosted antitumor efficacy, the acidity-responsive nanovehicles for doxorubicin (DOX) delivery were fabricated by tailor-made co-assembly of amphiphilic PEGylated chitosan20k and hydrophobic poly(lactic-co-glycolic acid) (PLGA) segments at pH 8.5. The attained DOX-loaded PEGylated chitosan20k/PLGA nanoparticles (DOX-PC20kPNs) were characterized to have a spherical shape composed of drug-encapsulated chitosan20k/PLGA-constituted solid core surrounded by hydrophilic PEG shells. Compared to non-pH-sensitive DOX-loaded PLGA nanoparticles (DOX-PNs), the DOX-PC20kPNs displayed outstanding colloidal stability under serum-containing condition and tended to swell in weak acidic milieu upon increased protonation of chitosan20k within hybrid cores, thus accelerating drug release. The in vitro cellular uptake and cytotoxicity studies revealed that the DOX-PC20kPNs after being endocytosed by prostate TRAMP-C1 cancer cells rapidly liberated drug, thus promoting drug accumulation in nuclei to enhance anticancer potency. Moreover, the hydrated PEG shells of DOX-PC20kPNs remarkably reduced their uptake by macrophage-like RAW264.7 cells. Importantly, in vivo animal findings showed that the DOX-PC20kPNs exhibited the capability of inhibiting TRAMP-C1 tumor growth superior to free hydrophobic DOX molecules and DOX-PNs, demonstrating the great potential in cancer chemotherapy.
Assuntos
Antineoplásicos , Quitosana , Nanopartículas , Neoplasias , Animais , Antineoplásicos/farmacologia , Quitosana/química , Doxorrubicina/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Concentração de Íons de Hidrogênio , Masculino , Nanopartículas/química , Polietilenoglicóis/químicaRESUMO
Liver cancer is a leading cause of cancerrelated mortality globally. Since hepatitis virus infections have been strongly associated with the incidence of liver cancer, studies concerning the effects of antiviral drugs on liver cancer have attracted great attention in recent years. The present study investigated the effects of two antihepatitis virus drugs, lamivudine and ribavirin, and one antiinfluenza virus drug, oseltamivir, on liver cancer cells to assess alternative methods for treating liver cancer. MTT assays, wound healing assays, Τranswell assays, flow cytometry, immunoblotting, ELISA, immunofluorescence staining and a xenograft animal model were adopted to verify the effects of lamivudine, ribavirin and oseltamivir on liver cancer cells. Treatment with ribavirin and oseltamivir for 24 and 48 h significantly decreased the viability of both Huh-7 and HepG2 cells compared with that of THLE3 cells in a dosedependent manner. The subsequent investigations focused on oseltamivir, considering the more serious clinical adverse effects of ribavirin than those of oseltamivir. Significantly decreased migration and invasion were observed in both Huh-7 and HepG2 cells that were treated with oseltamivir for 24 and 48 h. In addition, oseltamivir significantly increased autophagy in Huh7 cells, as revealed by the significantly higher ratios of LC3II/LC3I, increased expression of Beclin1, and decreased expression of p62, whereas no significant increases in the expression of apoptosisrelated proteins, including Apaf1, cleaved caspase3, and cleaved PARP1, were detected. Notably, apoptosis and autophagy were significantly increased in HepG2 cells in the presence of oseltamivir, as revealed by the significant increases in the expression of Apaf1, cleaved caspase3, and cleaved PARP1, the higher ratios of LC3II/LC3I, the increased expression of Beclin1, and the decreased expression of p62. Additionally, significant inhibitory effects of oseltamivir on xenografted Huh7 cells in athymic nude mice were observed. The present study, for the first time to the best of our knowledge, reported the differential effects of oseltamivir on inducing liver cancer cell death both in vitro and in vivo and may provide an alternative approach for treating liver cancer.
Assuntos
Antivirais/farmacologia , Autofagia , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Oseltamivir/farmacologia , Animais , Apoptose , Carcinoma Hepatocelular/patologia , Proliferação de Células , Feminino , Humanos , Neoplasias Hepáticas/patologia , Camundongos , Camundongos Nus , Invasividade Neoplásica , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
A unique region of human parvovirus B19 virusVP1 (B19VVP1u) has been linked to a variety of cardiac disorders. However, the precise role of B19VVP1u in inducing cardiac injury remains unknown. The present study investigated the effects of B19VVP1u and different regions of B19VVP1u, including B19VVP1uA (residues 160), B19VVP1uB (residues 61129), B19VVP1uC (residues 130195) and B19VVP1uD (residues 196227), on inducing cardiac injury in naïve mice by zymography, immunoblotting, H&E staining and cytokine immunoassay. A significantly higher MMP9/MMP2 ratio and increased levels of inflammatory cytokines, including IL6 and IL1ß, were detected in the left ventricles of the mice injected with B19Vnonstructural protein 1 (B19VNS1) and B19VVP1u, accompanied by increased expression levels of phosphorylated (p)ERK and pP38. Significantly upregulated expression levels of atrial natriuretic peptide (ANP), hearttype fatty acidbinding protein (HFABP) and creatine kinase isoenzymeMB (CKMB), which are wellknown cardiac injury markers, as well as increased infiltration of lymphocytes, were detected in the left ventricles of the mice injected with B19VVP1, B19VNS1 and B19VVP1u. Moreover, a significantly higher MMP9/MMP2 ratio and increased levels of IL6 and IL1ß were observed in the left ventricles of the mice injected with B19VVP1u, B19VVP1uA, B19VVP1uB and B19VVP1uC, accompanied by upregulated pERK and pP38 expression. Notably, significantly lower levels of IL6 and IL1ß were observed in the left ventricles of the mice injected with B19VVP1uD. Furthermore, significantly increased ANP, HFABP and CKMB expression levels were detected in the left ventricles of the mice injected with B19VVP1u, B19VVP1uA and B19VVP1uB, along with enhanced infiltration of lymphocytes. Significantly higher serum IL1ß, IL6, TNFα and IFNγ levels were also detected in the mice injected with B19VVP1u, B19VVP1uA and B19VVP1uB. To the best of our knowledge, the findings of the present study were the first to demonstrate that the Nterminal region (residues 1129) of B19VVP1u induces an increase in the levels of cardiac injury markers, thus providing evidence for understanding the possible functional regions within B19VVP1u.
Assuntos
Proteínas do Capsídeo/imunologia , Traumatismos Cardíacos/imunologia , Infecções por Parvoviridae/complicações , Parvovirus B19 Humano/imunologia , Animais , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/isolamento & purificação , Citocinas/sangue , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Traumatismos Cardíacos/sangue , Traumatismos Cardíacos/patologia , Traumatismos Cardíacos/virologia , Interações Hospedeiro-Patógeno/imunologia , Humanos , Camundongos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Transdução de Sinais/imunologiaRESUMO
INTRODUCTION: Glioblastoma multiforme (GBM) is the most aggressive glioma, and its diffuse nature makes resection of it difficult. Moreover, even with the administration of postoperative radiotherapy and chemotherapy, prolonged remission is often not achieved. Hence, innovative or alternative treatments for GBM are urgently required. Traditional Chinese herbs and their functional components have long been used in the treatment of various cancers, including GBM. The current study investigated the antitumor activity of Wedelia chinensis and its major functional components, luteolin and apigenin, on GBM. MATERIALS AND METHODS: MTT assay, Transwell migration assay, and flow cytometry analysis were adopted to assess the cell viability, invasive capability, and cell cycle. Immunofluorescence staining and Western blotting were used to detect the expressions of apoptotic and autophagy-related signaling molecules. RESULTS: The W. chinensis extract (WCE) significantly inhibited the proliferation and invasive ability of both GBM8401 and U-87MG cells in a dose-dependent manner. Moreover, differential effects of WCE on GBM8401 and U-87MG cells were observed: WCE induced apoptosis in GBM8401 cells and autophagy in U-87MG cells. Notably, WCE had significant effects in reducing the cell survival and invasive capability of both GBM8401 and U-87MG cells than the combination of luteolin and apigenin. CONCLUSIONS: Taken together, these findings indicate the potential of using WCE and the combination of luteolin and apigenin for GBM treatment. However, further investigations are warranted before considering recommending the clinical use of WCE or the combination of luteolin and apigenin as the standard for GBM treatment.