RESUMO
10-Hydroxycamptothecin (HCPT) is a widely used clinical anticancer drug but has a significant side effect profile. Melatonin has a beneficial impact on the chemotherapy of different cancer cells and reproductive processes, but the effect and underlying molecular mechanism of melatonin's involvement in the HCPT-induced side effects in cells, especially in the testicular cells, are poorly understood. In this study, we found that melatonin therapy significantly restored HCPT-induced testicular cell damage and did not affect the antitumor effect of HCPT. Further analysis found that melatonin therapy suppressed HCPT-induced DNA damage associated with ataxia-telangiectasia mutated- and Rad3-related and CHK1 phosphorylation levels in the testis. Changes in apoptosis-associated protein levels (Bax, Bcl-2, p53, and Cleaved caspase-3) and in reactive oxygen species-associated proteins (Nrf2 and Keap1) and index (malondialdehyde and glutathione) suggested that melatonin treatment relieved HCPT-induced cell apoptosis and oxidative damage, respectively. Mechanistically, melatonin-activated autophagy proteins (ATG7, Beclin1, and LC3bII/I) may induce p62-dependent autophagy to degrade Keap1, eliciting Nrf2 from Keap1-Nrf2 interaction to promote antioxidant enzyme expression such as HO-1, which would salvage HCPT-induced ROS production and mitochondrial dysfunction. Collectively, this study reveals that melatonin therapy may protect testicular cells from HCPT-induced damage via the activation of autophagy, which alleviates oxidative stress, mitochondrial dysfunction, and cell apoptosis.
Assuntos
Apoptose , Camptotecina , Melatonina , Estresse Oxidativo , Transdução de Sinais , Testículo , Animais , Masculino , Camundongos , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Camptotecina/farmacologia , Camptotecina/análogos & derivados , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Melatonina/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Proteína Sequestossoma-1/metabolismo , Transdução de Sinais/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/metabolismoRESUMO
Metal-organic frameworks (MOFs) are considered to be promising materials for drug delivery. In this work, a Zinc-based MOF nanocomposite IRMOF-3 was introduced as a drug carrier for 10-hydroxycamptothecine (HCPT). Without an extra drug-loading process, a nanoscale drug delivery material HCPT@IRMOF-3 was prepared via one-pot synthesis. The composition and structure of the material were investigated, and the drug release character was measured. Compared with preparing IRMOF-3 first and loading the drug, the one-pot-prepared HCPT@IRMOF-3 exhibited a higher drug-loading capacity. The material presented pH-responsive release. The HCPT release rate at pH 5.0 was significantly higher than that at pH 7.4. The cytotoxicity experiments showed that IRMOF-3 was non-toxic, and HCPT@IRMOF-3 exhibited notable cytotoxicity to Hela and SH-SY5Y cells. One-pot synthesis is a simple and rapid method for the preparation of an MOF drug delivery system, and IRMOF-3 can be potentially used in pH-responsive drug delivery systems.
Assuntos
Antineoplásicos , Nanopartículas , Neuroblastoma , Humanos , Antineoplásicos/farmacologia , Antineoplásicos/química , Sistemas de Liberação de Medicamentos/métodos , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Nanopartículas/química , Concentração de Íons de HidrogênioRESUMO
CDKL1 is a cyclin-dependent kinase-like kinase that is highly expressed in diverse types of cancer cells. However, the role of CDKL1 in the chemoresistance of oral squamous cell carcinoma (OSCC) remains largely undefined. Here, we explored the role of CDKL1 in the chemoresistance of the human OSCC cell line CAL27 to hydroxycamptothecin (HCPT). Real-time quantitative polymerase chain reaction and western blotting revealed that exposure of CAL27â¯cells to HCPT led to a marked increase in the expression of CDKL1 at the mRNA and protein levels. Knockdown of CDKL1 significantly suppressed cell proliferation and induced cell cycle G0/G1 phase arrest in CAL27â¯cells based on the results of MTT and flow cytometry assays, respectively. CAL27 cells displayed attenuated biological activity of the cell population. After treatment with HCPT, whereas CDKL1 overexpression increased the resistance to HCPT of the remaining cells. Moreover, the western blot showed that the expression of cleaved-caspase 3 and phosphorylated ataxia telangiectasia mutated proteins was upregulated by HCPT treatment in CAL27â¯cells. Furthermore, CDKL1 overexpression partially reversed the inhibitory effects of HCPT in CAL27â¯cells. These results suggest that CDKL1 overexpression decreased the chemosensitivity of OSCC cells to HCPT, indicating a potential strategic approach for reversing the HCPT resistance in human OSCC.
Assuntos
Camptotecina/análogos & derivados , Carcinoma de Células Escamosas/tratamento farmacológico , Quinases Ciclina-Dependentes/metabolismo , Resistencia a Medicamentos Antineoplásicos , Neoplasias Bucais/tratamento farmacológico , Proteínas do Tecido Nervoso/metabolismo , Apoptose/efeitos dos fármacos , Camptotecina/farmacologia , Camptotecina/uso terapêutico , Carcinoma de Células Escamosas/patologia , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Neoplasias Bucais/patologia , RNA Interferente Pequeno/metabolismoRESUMO
In this paper, shape regulated anticancer activities as well as systematic toxicities of hydroxycamptothecin nanorods and nanoparticles (HCPT NRs and NPs) were systematically studied. In vitro and in vivo therapeutic efficacies were evaluated in cancer cells and tumor-bearing mice, indicating that NRs possessed superior antitumor efficacy over NPs at the equivalent dose, while systematic toxicity of the differently shaped nanodrugs assessed in healthy mice, including the maximum tolerated dose, blood analysis and histology examinations and so on, suggested that the NRs also caused higher toxicities than NPs, and also had a long-term toxicity. These results imply that the balance between anticancer efficiency and systematic toxicity of drug nanocrystals should be fully considered in practice, which will provide new concept in the future design of drug nanocrystals for cancer therapy. From the Clinical Editor: Advances in nanotechnology have enabled the design of novel nanosized drugs for the treatment of cancer. One of the interesting findings thus far is the different biological effects seen with different shaped nanoparticles. In this article, the authors investigated and compared the anticancer activities of hydroxycamptothecin nanorods and nanoparticles. The experimental data would provide a better understanding for future drug design.
Assuntos
Camptotecina/análogos & derivados , Nanopartículas/administração & dosagem , Nanopartículas/ultraestrutura , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/patologia , Animais , Camptotecina/administração & dosagem , Camptotecina/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cristalização/métodos , Composição de Medicamentos/métodos , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Tamanho da Partícula , Resultado do TratamentoRESUMO
10-Hydroxycamptothecin (HCPT), a plant alkaloid isolated from Camptotheca acuminate, is known as a planted-derived insecticide, however, the specific mechanism in insect cells is still unclear. In this study, we treated the ovarian cell line of the silkworm, BmN-SWU1, with different HCPT doses for durations ranging from 0 to 72h. The apoptosis morphology was evident after 72h of incubation and included cell protuberance, concentrated cytoplasm and apoptotic bodies. We observed DNA fragmentation and cell apoptosis after HCPT treatment. The disruption of mitochondrial distribution, activation of the intracellular mitochondrial permeability transition pore, and release of cytochrome c during HCPT-induced apoptosis in dose and time-dependent manner indicate the involvement of mitochondria in BmN-SWU1 cells. Caspase-9 and -3 activities increased gradually with the duration of incubation time. In conclusion, HCPT has a significant effect to initiate the intrinsic mitochondrial pathway in silkworm cells, providing a theoretical basis for better application of plant-derived insecticide in pest control.
Assuntos
Camptotecina/análogos & derivados , Mitocôndrias/efeitos dos fármacos , Animais , Apoptose , Bombyx , Camptotecina/farmacologia , Linhagem Celular , Feminino , Ovário/citologiaRESUMO
Bcl-2 family proteins have been reported previously to play important roles in the mitochondrial apoptotic pathway. Particularly, Bmbuffy has been identified as a key homologue of Bcl-2 in silkworm; however, its exact function is unknown. In this study, we investigated the role of Bmbuffy in hydroxycamptothecine (HCPT)-induced apoptosis of BmN-SWU1 cells. By conducting confocal microscopy studies, we found that Bmbuffy is located on the outer membrane of mitochondria and endoplasmic reticulum (ER). Furthermore, we discovered that the hydrophobic transmembrane domain at the COOH terminus is a putative anchor for the subcellular localization of Bmbuffy. Overexpression of Bmbuffy inhibited cytochrome c release, activation of caspase-3 and cell apoptosis, while RNAi-mediated silencing of Bmbuffy promoted apoptosis. In the absence of a hydrophobic membrane anchor, we revealed that Bmbuffy is unable to block apoptosis. These results indicate that Bmbuffy acts as an anti-apoptotic protein, located on the mitochondrial outer membrane and is involved in the mitochondrial apoptotic pathway. Moreover, in HCPT-induced apoptosis, we showed that the translocation of endogenous Bmp53 from the nucleus to the mitochondria is a slow and progressive process, followed by cytochrome c release. This suggests that mitochondrial Bmp53 accumulation may contribute to membrane permeability. The co-localization of Bmp53 and Bmbuffy suggests the interaction of the two proteins, which was further confirmed by Co-IP assay. In addition, overexpression of Bmp53 increased cytochrome c release and the cell apoptotic rate, whereas Bmbuffy overexpression blocked these. All the data suggest that Bmbuffy functions as an anti-apoptotic protein and interacts with Bmp53 in HCPT-induced apoptosis of silkworm cells.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose , Bombyx/metabolismo , Proteínas de Insetos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Apoptose/efeitos dos fármacos , Camptotecina/análogos & derivados , Camptotecina/farmacologia , Caspase 3/metabolismo , Linhagem Celular , Citocromos c/metabolismo , Retículo Endoplasmático/metabolismo , Membranas Mitocondriais/metabolismo , Permeabilidade , Proteína Supressora de Tumor p53/metabolismoRESUMO
Mycorrhizal associations are known to improve the hydro-mineral nutrition of their host plants. However, the importance of mycorrhizal symbiosis for plant potassium nutrition has so far been poorly studied. We therefore investigated the impact of the ectomycorrhizal fungus Hebeloma cylindrosporum on the potassium nutrition of Pinus pinaster and examined the involvement of the fungal potassium transporter HcTrk1. HcTrk1 transcripts and proteins were localized in ectomycorrhizas using in situ hybridization and EGFP translational fusion constructs. Importantly, an overexpression strategy was performed on a H. cylindrosporum endogenous gene in order to dissect the role of this transporter. The potassium nutrition of mycorrhizal pine plants was significantly improved under potassium-limiting conditions. Fungal strains overexpressing HcTrk1 reduced the translocation of potassium and phosphorus from the roots to the shoots of inoculated plants in mycorrhizal experiments. Furthermore, expression of HcTrk1 and the phosphate transporter HcPT1.1 were reciprocally linked to the external inorganic phosphate and potassium availability. The development of these approaches provides a deeper insight into the role of ectomycorrhizal symbiosis on host plant K(+) nutrition and in particular, the K(+) transporter HcTrk1. The work augments our knowledge of the link between potassium and phosphorus nutrition via the mycorrhizal pathway.
Assuntos
Proteínas Fúngicas/metabolismo , Hebeloma/metabolismo , Interações Hospedeiro-Patógeno , Micorrizas/metabolismo , Fósforo/metabolismo , Pinus/metabolismo , Potássio/metabolismo , Transporte Biológico/efeitos dos fármacos , DNA Bacteriano/genética , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Hebeloma/efeitos dos fármacos , Hebeloma/genética , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Hifas/efeitos dos fármacos , Hifas/metabolismo , Micorrizas/efeitos dos fármacos , Micorrizas/genética , Fenótipo , Fósforo/farmacologia , Pinus/efeitos dos fármacos , Pinus/microbiologia , Potássio/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Plântula/efeitos dos fármacos , Plântula/metabolismo , Plântula/microbiologia , Sódio/metabolismoRESUMO
Mycorrhizal exchange of nutrients between fungi and host plants involves a specialization and polarization of the fungal plasma membrane adapted for the uptake from the soil and for secretion of nutrient ions towards root cells. In addition to the current progress in identification of membrane transport systems of both symbiotic partners, data concerning the transcriptional and translational regulation of these proteins are needed to elucidate their role for symbiotic functions. To answer whether the formerly described Pi-dependent expression of the phosphate transporter HcPT1.1 from Hebeloma cylindrosporum is the result of its promoter activity, we introduced promoter-EGFP fusion constructs in the fungus by Agrotransformation. Indeed, HcPT1.1 expression in pure fungal cultures quantified and visualized by EGFP under control of the HcPT1.1 promoter was dependent on external Pi concentrations, low Pi stimulating the expression. Furthermore, to study expression and localization of the phosphate transporter HcPT1.1 in symbiotic conditions, presence of transcripts and proteins was analyzed by the in situ hybridization technique as well as by immunostaining of proteins. In ectomycorrhiza, expression of the phosphate transporter was clearly enhanced by Pi-shortage indicating its role in Pi nutrition in the symbiotic association. Transcripts were detected in external hyphae and in the hyphal mantle, proteins in addition also within the Hartig net. Exploiting the transformable fungus H. cylindrosporum, Pi-dependent expression of the fungal transporter HcPT1.1 as result from its promoter activity as well as transcript and protein localization in ectomycorrhizal symbiosis are shown.
Assuntos
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Micorrizas/metabolismo , Proteínas de Transporte de Fosfato/genética , Proteínas de Transporte de Fosfato/metabolismo , Fosfatos/metabolismo , Regiões Promotoras Genéticas , Hebeloma/genética , Hebeloma/metabolismo , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Micorrizas/genética , Micorrizas/crescimento & desenvolvimento , Pinus/microbiologia , Pinus/fisiologia , Transporte Proteico , SimbioseRESUMO
Corneal fibrosis is a common outcome of inappropriate repair associated with trauma or ocular infection. Altered biomechanical properties with increased corneal stiffness is a feature of fibrosis that cause corneal opacities, resulting in severe visual impairment and even blindness. The present study aims to determine the effect of hydroxycamptothecin (HCPT) and matrix stiffness on transforming growth factor-ß1 (TGF-ß1)-induced fibrotic processes in human corneal fibroblasts (HTK cells). HTK cells were cultured on substrates with different stiffnesses ("soft", â¼261 kPa; "stiff", â¼2.5 × 103 kPa) and on tissue culture plastic (TCP, â¼106 kPa) and simultaneously treated with or without 1 µg/mL HCPT and 10 ng/mL TGF-ß1. We found that HCPT induced decreased cell viability and antiproliferative effects on HTK cells. TGF-ß1-induced expression of fibrosis-related genes (FN1, ACTA2) was reduced if the cells were simultaneously treated with HCPT. Substrate stiffness did not affect the expression of fibrosis-related genes. The TGF-ß1 induced expression of FN1 on both soft and stiff substrates was reduced if cells were simultaneously treated with HCPT. However, this trend was not seen for ACTA2, i.e., the TGF-ß1 induced expression of ACTA2 was not reduced by simultaneous treatment of HCPT in either soft or stiff substrate. Instead, HCPT treatment in the presence of TGF-ß1 resulted in increased gene expression of keratocyte phenotype makers (LUM, KERA, AQP1, CHTS6) on both substrate stiffnesses. In addition, the protein expression of keratocyte phenotype makers LUM and ALDH3 was increased in HTK cells simultaneously treated with TGF-ß1 and HCPT on stiff substrate as compared to control, i.e., without HCPT. In conclusion, we found that HCPT can reduce TGF-ß1-induced fibrosis and promote the keratocyte phenotype in a substrate stiffness dependent manner. Thus, HCPT stimulation might be an approach to stimulate keratocytes in the appropriate healing stage to avoid or reverse fibrosis and achieve more optimal corneal wound healing.
Assuntos
Fibroblastos , Fator de Crescimento Transformador beta1 , Humanos , Fator de Crescimento Transformador beta1/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Células Cultivadas , Fibroblastos/metabolismo , Fibroblastos/patologia , FibroseRESUMO
Head and neck squamous carcinoma (HNSCC) is the seventh most common cancer worldwide. Targeted therapeutic drugs for HNSCC are still being explored. Among them, (S)-10-hydroxycamptothecin (10-HCPT), a specific inhibitor of TOP1, functions by DNA double-strand breaks that can inhibit DNA replication and trigger apoptotic cell death subsequently. Previous studies have reported that MLN4924 exerts potent anti-tumor effects by inhibiting cullin-RING ligases and causing substrate accumulation in a variety of cancers. Here, we show that MLN4924 effectively causes dose-dependent accumulation of topoisomerase I (TOP1) and blocks TOP1 ubiquitination. Importantly, neddylation inhibition with MLN4924 acts synergistically with 10-HCPT to suppress cell growth, migration and apoptosis in HNSCC cells. Mechanistically, transcriptome sequencing shows that the cytotoxic effects of the combination of MLN4924 and 10-HCPT may involve activation of the NFKB1 pathway. Taken together, our results suggest that combined treatment with MLN4924 and 10-HCPT may be an effective strategy in HNSCC.
Assuntos
Camptotecina , Neoplasias de Cabeça e Pescoço , Humanos , Carcinoma de Células Escamosas de Cabeça e Pescoço/tratamento farmacológico , Camptotecina/farmacologia , Neoplasias de Cabeça e Pescoço/tratamento farmacológicoRESUMO
Pseudorabies virus (PRV), a member of the subfamily alphaherpesvirinae, is one of the most important pathogenes that cause acute death in infected pigs and leads to substantial economic losses in the global swine industry. Recently, China's emerging PRV mutant strains resulted in the traditionally commercial vaccines not providing complete protection. Some studies reported that PRV could infect humans and cause endophthalmitis and encephalitis under certain circumstances. It is necessary to develop alternative manners to control the virus infection. Here, by screening a library of natural products, (S)-10-Hydroxycamptothecin (10-HCPT) was revealed to inhibit PRV replication with a selective index of 270.04. And 10-HCPT inhibited PRV replication by blocking the viral genome replication but not inhibiting the viral attachment, internalization, and release. RNA interference assay showed that 10-HCPT inhibited PRV replication by targeting DNA topoisomerase 1 (TOP1). Meanwhile, 10-HCPT treatment induced DNA damage response and stimulated antiviral innate immunity. Animal challenge experiments showed that 10-HCPT effectively alleviated clinical signs and hispathology, and increased INF-ß responses in lung and brain tissues of mice induced by PRV infection. The results demonstrate that 10-HCPT is a promising therapeutic agent to control PRV infection.
Assuntos
Produtos Biológicos , Herpesvirus Suídeo 1 , Pseudorraiva , Doenças dos Roedores , Doenças dos Suínos , Animais , Antivirais/farmacologia , Antivirais/uso terapêutico , Produtos Biológicos/farmacologia , Camptotecina/análogos & derivados , Linhagem Celular , Proliferação de Células , Dano ao DNA , Herpesvirus Suídeo 1/genética , Imunidade Inata , Camundongos , Vacinas contra Pseudorraiva , SuínosRESUMO
The combination of chemotherapy and immunotherapy motivates a potent immune system by triggering immunogenic cell death (ICD), showing great potential in inhibiting tumor growth and improving the immunosuppressive tumor microenvironment (ITM). However, the therapeutic effectiveness has been restricted by inferior drug bioavailability. Herein, we reported a universal bioresponsive doxorubicin (DOX)-based nanogel to achieve tumor-specific co-delivery of drugs. DOX-based mannose nanogels (DM NGs) was designed and choosed as an example to elucidate the mechanism of combined chemo-immunotherapy. As expected, the DM NGs exhibited prominent micellar stability, selective drug release and prolonged survival time, benefited from the enhanced tumor permeability and prolonged blood circulation. We discovered that the DOX delivered by DM NGs could induce powerful anti-tumor immune response facilitated by promoting ICD. Meanwhile, the released mannose from DM NGs was proved as a powerful and synergetic treatment for breast cancer in vitro and in vivo, via damaging the glucose metabolism in glycolysis and the tricarboxylic acid cycle. Overall, the regulation of tumor microenvironment with DOX-based nanogel is expected to be an effectual candidate strategy to overcome the current limitations of ICD-based immunotherapy, offering a paradigm for the exploitation of immunomodulatory nanomedicines.
RESUMO
Nanoparticulate drug delivery systems (Nano-DDSs) have emerged as possible solution to the obstacles of anticancer drug delivery. However, the clinical outcomes and translation are restricted by several drawbacks, such as low drug loading, premature drug leakage and carrier-related toxicity. Recently, pure drug nano-assemblies (PDNAs), fabricated by the self-assembly or co-assembly of pure drug molecules, have attracted considerable attention. Their facile and reproducible preparation technique helps to remove the bottleneck of nanomedicines including quality control, scale-up production and clinical translation. Acting as both carriers and cargos, the carrier-free PDNAs have an ultra-high or even 100% drug loading. In addition, combination therapies based on PDNAs could possibly address the most intractable problems in cancer treatment, such as tumor metastasis and drug resistance. In the present review, the latest development of PDNAs for cancer treatment is overviewed. First, PDNAs are classified according to the composition of drug molecules, and the assembly mechanisms are discussed. Furthermore, the co-delivery of PDNAs for combination therapies is summarized, with special focus on the improvement of therapeutic outcomes. Finally, future prospects and challenges of PDNAs for efficient cancer therapy are spotlighted.
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The poor water solubility and severe side effects of hydroxycamptothecin (HCPT) limit its clinical application; therefore, it is necessary to synthesize applicable nanodrug carriers with good solubility to expand the applications of HCPT. In this study, a hydrophilic metal-organic framework (MOF) with amphoteric carboxylic acid ligands as linkers was first synthesized and characterized. Then, water-soluble acrylamide and methacrylic acid were applied as monomers to prepare a water-soluble polyacid polymer MOF@P, which had a solubility of 370 µg/mL. The effects of the MOF@P material on the HCPT loading and solubility were investigated. The results showed that the polymer material could improve the HCPT solubility in water. Moreover, the in vitro release study indicated that the MOF@P polymeric composite exhibited a sustained-release effect on HCPT, with a cumulative release rate of 30.18% in 72 h at pH 7.4. Furthermore, the cytotoxicity test demonstrated that the hydrophilic MOF and the MOF@P had low cell toxicities. The results indicate that the prepared MOF@P polymeric complex can be applied for the sustained release of HCPT in clinics.
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Tumor multidrug resistance (MDR) is one of the main reasons for the failure of clinical chemotherapy. Here, a bio-responsive anti-drug-resistant polymer micelle that can respond to the reductive GSH in the tumor microenvironment (TME) for delivery of HCPT was designed. A new type of polymer with anti-drug resistance and anti-tumor effect was synthesized and used to encapsulated HCPT to form reduction-sensitive micelles (PDSAH) by a thin-film dispersion method. It is demonstrated that the micelle formulation improves the anti-tumor activity and biosafety of HCPT, and also plays a significant role in reversing the drug resistance, which contributes to inhibiting the tumor growth and prolonging the survival time of H22 tumor-bearing mice. The results indicate that this nanoplatform can serve as a flexible and powerful system for delivery of other drugs that are tolerated by tumors or bacteria.
Assuntos
Camptotecina , Micelas , Animais , Camptotecina/análogos & derivados , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Camundongos , PolímerosRESUMO
10-HCPT is a topoisomerase I inhibitor effective in the treatment of liver cancer but its use is hampered by its resistance. The expression of hypoxia-inducible factor-1α (HIF-1α) is reportedly upregulated in liver cancer tissues, which is directly linked to the resistance of 10-HCPT. While BBR can significantly decrease the level of HIF-1α according to the literature report. Thus, the aim of this study was to prepare a novel intravenous 10-HCPT-BBR-loaded lipid microsphere (LM) and evaluate their synergistic effect on liver cancer treatment. The optimal preparation mainly included 10.0% oil phase (medium-chain triglyceride:long-chain triglyceride = 1:1), emulsifier (egg lecithin E80 and pluronic F68), antioxidant (0.02% NaHSO3), and pH regulator (0.1 mol/L Hcl). Then, the behaviors of BBR-10-HCPT loaded LM in vitro and in vivo were systematically investigated. In vitro, it showed an obvious sustained-release effect in different release mediums, good physicochemical stability at accelerated and long-term storage conditions, and great anti-proliferative capability toward human liver cancer Hep-3B cells. In vivo, the prepared LM exhibited a longer half-life and higher AUC compared to BBR injection and 10-HCPT injection. More importantly, it was found that The LM was distributed more in the liver, spleen, and tumors, but less in the lungs and heart, especially in the lung. And then, it showed significant inhibition of tumor growth against nude mouse with Hep-3B tumor, and the tumor inhibition rate reached 91.55%. Thus, the data obtained in our study suggested that BBR combined with 10-HCPT can raise curative effect and reduce the toxicity of 10-HCPT.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Berberina/farmacologia , Camptotecina/análogos & derivados , Carcinoma Hepatocelular , Neoplasias Hepáticas , Microesferas , Inibidores da Topoisomerase I/farmacologia , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Berberina/administração & dosagem , Berberina/química , Camptotecina/administração & dosagem , Camptotecina/química , Camptotecina/farmacologia , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Camundongos , Camundongos Nus , Ratos , Inibidores da Topoisomerase I/administração & dosagem , Inibidores da Topoisomerase I/química , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: This study aims to study the preparation method of arsenic trioxide (As2O3) polylactic-co-glyconlic acid (PLGA) microspheres and 10-hydroxycamptothecin (HCPT) PLGA microspheres and explore their therapeutic effects as embolic agents for VX2 hepatocellular carcinoma in rabbits. METHODS: As2O3 and HCPT PLGA microspheres were prepared by multiple emulsion solvent evaporation method. Scanning electron microscopy (SEM) and particle size distribution were used to analyze the morphology, the drug sustained release ability was observed by the release of microspheres in vitro. The rabbit model of VX2 hepatocellular carcinoma was established and the hepatocellular carcinoma was treated with combined microspheres. The therapeutic effects were detected by qPCR, western blotting, HE staining and immunohistochemical methods. RESULTS: The PLGA microspheres loaded with As2O3 and HCPT were successfully prepared by optimizing the ratio. The particle size was between 30 and 50 µm. In vitro release results showed that PLGA microspheres loaded with As2O3 released completely in 10 days and PLGA microspheres loaded with HCPT released completely in 12 days. Western blotting and qPCR results showed that the expression of ALDH1A1 and Nanog decreased significantly in treatment group. HE staining and immunohistochemical analysis showed that the expression of CD31, HIF and VEGF decreased significantly and the apoptosis of tissues was obvious. CONCLUSION: The combination of As2O3 and HCPT PLGA microspheres as embolization for VX2 hepatocellular carcinoma in rabbits has significant therapeutic effect.
Assuntos
Antineoplásicos/administração & dosagem , Trióxido de Arsênio/administração & dosagem , Camptotecina/análogos & derivados , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Microesferas , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Animais , Camptotecina/administração & dosagem , Modelos Animais de Doenças , CoelhosRESUMO
Nanoparticles decorated with hydrophilic PEG chains had been emerged as effective drug delivery system due to their biocompatibility and biodegradability, while, the influence of their architecture on antitumor efficacy remained challenging. In this study, the linear poly(ethylene glycol) (PEG45), brush oligo(triethylene glycol) (TEG10), and oligo(ethylene glycol) dendron (G2), which showed the similar molar mass but different architecture, were utilized as nanocarriers to prepare 10-hydroxycamptothecin (HCPT) nanoparticles. It was found that all of three HCPT NPs (PEG45 NPs, TEG10 NPs, and G2 NPs) presented similar DLCs (â¼60%), zeta potentials (-13 to -16â¯mv), and stabilities, meanwhile, the particle sizes, morphologies, release profiles, and anticancer efficacies were affected by the architecture of nanocarriers. Changing the architecture from linear, brush to dendron, the mean particles diameter was decreased from 240 to 170â¯nm, the sustained releases were elongated from 5 to 9â¯days. More importantly, the cytotoxicity of G2 NPs based on OEG dendron was enhanced significant comparing with linear PEG45 NPs, the IC50 was decreased almost 10.1-fold (pâ¯<â¯0.01). Besides, the tumor inhibition rate of G2 NPs was 1.7-fold higher than PEG45 NPs, showing significantly optimized antitumor efficacy in vivo. These results suggested the architecture of nanocarriers could affect the antitumor activity, due to the steric hindrance of nanocarriers influence the morphologies of HCPT-loaded nanoparticles.
Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Camptotecina/análogos & derivados , Portadores de Fármacos/química , Etilenoglicol/química , Neoplasias/tratamento farmacológico , Animais , Antineoplásicos Fitogênicos/farmacocinética , Camptotecina/administração & dosagem , Camptotecina/farmacocinética , Linhagem Celular Tumoral/transplante , Modelos Animais de Doenças , Composição de Medicamentos/métodos , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Humanos , Injeções Intravenosas , Camundongos , Camundongos Endogâmicos BALB C , Conformação Molecular , Nanopartículas/química , Tamanho da Partícula , Ratos , Distribuição TecidualRESUMO
Since the hydrophobic group is always essential to the synthesis of the drug-loaded nanoparticles, a majority of the methods rely heavily on organic solvent, which may not be completely removed and might be a potential threat to the patients. In this study, we completely "green" synthesized 10-hydroxycamptothecine (HCPT) loaded, folate (FA)-modified nanoneedles (HFNDs) for highly efficient cancer therapy with high drug loading, targeting property, and imaging capability. It should be noted that no organic solvent was used in the preparation process. In vitro cell uptake study and the in vivo distribution study showed that the HFNDs, with FA on the surface, revealed an obviously targeting property and entered the HeLa cells easier than the chitosan-HCPT nanoneedles without FA modified (NDs). The cytotoxicity tests illustrated that the HFNDs possessed better killing ability to HeLa cells than the individual drug or the NDs in the same dose, indicating its good anticancer effect. The in vivo anticancer experiment further revealed the pronounced anticancer effects and the lower side effects of the HFNDs. This new method without organic solvent will lead to a promising sustained drug delivery system for cancer diagnosis and treatment.
RESUMO
A redox-responsive supramolecular hydrogel system was developed for delivering 10-hydroxy camptothecin (HCPT). The hydrogel was formed by cleaving disulfide bond. The combination of hydrophobic HCPT with hydrogel was a simple and effective way to improve the solubility of HCPT and the drug loading capacity of delivery system. The transmission electron microscopy (TEM) image revealed the self-assembled hydrogel was long and thin nanofibers with a width of <10nm. Rheological test verified the hydrogel had fine physical properties. In vitro release experiment showed that the accumulative releasing percentages within 72h of HCPT-peptide hydrogels at 3.0%, 4.0%, 5.0% were 16.8%, 21.3%, and 26.8% respectively, which indicated the HCPT-peptide hydrogels had a significantly sustained-release characteristic. Besides, in vitro anticancer assay showed that HCPT-peptide hydrogels possessed a favorable anticancer efficacy. These results indicated that HCPT-peptide hydrogel had great potential for cancer treatment as a novel injectable drug delivery system.