RESUMO
MicroRNAs (miRNAs) can function as negative regulators of gene expression by binding to the 3'-untranslated region (3'-UTR) of target genes. The aberrant expression of miRNAs in neoplasm is extensively associated with tumorigenesis and cancer progression, including esophageal squamous cell carcinoma (ESCC). Our previous investigation has identified the oncogenic roles of Peroxiredoxin2 (PRDX2) in ESCC progression; however, its upstream regulatory mechanism remains to be elucidated. By merging the prediction results from miRWalk2.0 and miRNA differential expression analysis results based on The Cancer Genome Atlas Esophageal Carcinoma (TCGA-ESCA) database, eight miRNA candidates were predicted to be the potential regulatory miRNAs of PRDX2, followed by further identification of miR-92a-2-5p as the putative miRNA of PRDX2. Subsequent functional studies demonstrated that miR-92a-2-5p can suppress ESCC cell proliferation and migration, as well as tumor growth in subcutaneous tumor xenograft models, which might be mediated by the suppression of AKT/mTOR and Wnt3a/ß-catenin signaling pathways upon miR-92a-2-5p mimic transfection condition. These data revealed the tumor suppressive functions of miR-92a-2-5p in ESCC by targeting PRDX2.
Assuntos
Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , MicroRNAs , Humanos , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/patologia , Regulação Neoplásica da Expressão Gênica/genética , MicroRNAs/metabolismo , Peroxirredoxinas/genética , Peroxirredoxinas/metabolismo , AnimaisRESUMO
BACKGROUND: Disulfidptosis is a recently discovered programmed cell death pathway. However, the exact molecular mechanism of disulfidptosis in cutaneous melanoma remains unclear. METHODS: In this study, clustering analysis was performed using data from public databases to construct a prognostic model, which was subsequently externally validated. The biological functions of the model genes were then investigated through various experimental techniques, including qRT-PCR, Western blotting, CCK-8 assay, wound healing assay, and Transwell assay. RESULTS: We constructed a signature using cutaneous melanoma (CM) data, which accurately predicts the overall survival (OS) of patients. The predictive value of this signature for prognosis and immune therapy response was validated using multiple external datasets. High-risk CM subgroups may exhibit decreased survival rates, alterations in the tumor microenvironment (TME), and increased tumor mutation burden. We initially verified the expression levels of five optimum disulfidptosis-related genes (ODRGs) in normal tissues and CM. The expression levels of these genes were further confirmed in HaCaT cells and three melanoma cell lines using qPCR and protein blotting analysis. HLA-DQA1 emerged as the gene with the highest regression coefficient in our risk model, highlighting its role in CM. Mechanistically, HLA-DQA1 demonstrated the ability to suppress CM cell growth, proliferation, and migration. CONCLUSION: In this study, a novel signature related to disulfidptosis was constructed, which accurately predicts the survival rate and treatment sensitivity of CM patients. Additionally, HLA-DQA1 is expected to be a feasible therapeutic target for effective clinical treatment of CM.
Assuntos
Melanoma , Neoplasias Cutâneas , Humanos , Melanoma/genética , Melanoma/terapia , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/terapia , Imunoterapia , Aprendizado de Máquina , Microambiente Tumoral/genética , Melanoma Maligno CutâneoRESUMO
A facile and ingenious method to chemical etching-coordinating a metal-organic framework (MOF) followed by an annealing treatment was proposed to prepare Co3O4 nanoparticles uniformly dispersed in rational porous carbon nano-boxes (Co3O4@CNBs), which was further used to detect H2O2 released from living cells. The Co3O4@CNBs H2O2 sensor delivers much higher sensitivity than non-etching/coordinating Co3O4, offering a limit of detection of 2.32 nM. The wide working range covers 10 nM-359 µM H2O2, while possessing good selectivity and excellent reproducibility. Moreover, this biosensor was used to successfully real-time detect H2O2 released from living cells, including both healthy and tumor cells. The excellent performance holds great promise for Co3O4@CNBs's applications in electrochemical biomimetic sensing, particularly real-time monitor H2O2 released from living cells.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Biossensoriais/métodos , Carbono , Cobalto , Técnicas Eletroquímicas/métodos , Peróxido de Hidrogênio , Óxidos , Porosidade , Reprodutibilidade dos TestesRESUMO
In this work, we report a novel cell surface glycan analysis method based on persistent luminescence nanoparticle (PLNP) ZnGa2O4: Cr3+ (ZGC) as an optical probe. ZGC was first silanized by (3-Aminopropyl) triethoxysilane (APTES), followed by PEGylation with NHS-P EG-Biotin, which not only introduces biotin, but significantly improves the dispersibility and stability of the nanoparticles. Neutral-avidin was then coupled on ZGC surface through the specific biotin-avidin interaction, producing a ZGC-PEG-avidin nanoprobe. As for cell surface glycan detection, different surface glycans are recognized with their corresponding biotinylated lectins, which are then traced by ZGC-PEG-avidin. The persistent luminescence signal is recorded by a microtiter plate reader in time-resolved fluorescence mode. Glycans expression profiling on prostate cancer cell DU145 and normal prostate cell RWPE-1 was analyzed by the proposed detection platform. Similar results were observed from the conventional horseradish peroxidase (HRP)-catalyzed absorbent assay and confocal microscope-based fluorescence imaging, demonstrating the applicability of the proposed platform. The approach based on the long afterglow property of ZGC efficiently eliminates the background noise from cells and substrate, resulting in the best signal-to-noise ratio and high detection sensitivity.
Assuntos
Luminescência , Substâncias Luminescentes/química , Nanopartículas/química , Imagem Óptica , Polissacarídeos/análise , Células Cultivadas , HumanosRESUMO
Although a variety of approaches have been developed to analyze protein O-GlcNAcylation, efficient investigations on O-GlcNAcylation of proteins of interest in high-throughput manner are still in high demand to further explore its functionality. In this work, we first develop a powerful microarray platform for a sensitive, specific and high-throughput analysis of protein O-GlcNAcylation. The developed array biochip is then utilized to parallelly analyze the O-GlcNAcylation of three oncogenic transcription factors C-Myc, NF-κB and p53 in normal prostate epithelial cell (RWPE-1) and prostate cancer cell line (PC-3). The levels of O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) are also monitored by the microarray platform. The experimental results show that the overall O-GlcNAcylation and OGT expression level are obviously elevated in PC-3 as compared to RWPE-1. The protein expression-normalized O-GlcNAcylation of C-Myc and NF-κB in PC-3 is significantly higher than that in RWPE-1, while opposite result is observed from p53. In addition, the biological behaviors including proliferation and migration of PC-3â¯cells are also studied when OGA inhibitor Thiamet G is applied to elevate the total O-GlcNAcylation level.
Assuntos
Dispositivos Lab-On-A-Chip , N-Acetilglucosaminiltransferases/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Neoplasias da Próstata/metabolismo , Acilação , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , Masculino , NF-kappa B/metabolismo , Neoplasias da Próstata/patologia , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
A high-resolution colorimetric immunoassay platform has been developed based on enzyme-catalyzed multicolor generation and smartphone-assisted signal readout. The multi-color generation is accomplished in this system through the urease-catalyzed urea hydrolysis-induced color change of the pH indicator phenol red, from yellow to orange to red over pH 6.6 to 8.0. The color change is easily tailored by controlling the urease activity via the inhibitor silver ion (Ag+), the amount of which is in turn adjusted by alkaline phosphatase (ALP)-catalyzed ascorbic acid (AA) production. An ALP-linked colorimetric immunoassay is readily realized based on the above urease catalyzed multicolor generation system. Under optimal conditions, a limit of detection (LOD) of 1.73 ng mL-1 and a dynamic range from 0 to 18 ng mL-1 are achieved with rabbit IgG as a model analyte. A colored picture for each test is directly taken using a smartphone and then quantitatively analyzed with the free software ImageJ, eliminating the use of expensive and desktop equipment. The dose-dependent multicolor display is easier to distinguish with the naked eye for qualitative or semiquantitative detection over the traditional one-color system. The developed immunoassay scheme provides a promising platform for on-site testing or applications in resource-poor areas.
Assuntos
Colorimetria , Imunoensaio , Smartphone , Fosfatase Alcalina/química , Animais , Ácido Ascórbico/química , Limite de Detecção , Coelhos , Prata , Ureia/química , Urease/químicaRESUMO
FePO4 is biocompatible and can act as a kind of promising enzyme mimetic. Unfortunately, the electrical conductivity of FePO4 is poor. In order to enhance its conductivity, FePO4 was embedded into nanofibers consisting of amorphous carbon and reduced graphene oxide (rGO) by an electrospinning technique. As a sensing material for monitoring superoxide anion (O2â¢-) and typically operated at 0.5 V (vs. SCE), it displays high sensitivity (9.6 µAâ µM-1â cm-2), a low detection limit (9.7 nM at S/N = 3), a wide linear response range (10 nM to 10 µM), and fast response (1.6 s). Due to its low detection limit and fast response, the sensor in our perception has a large potential for detecting superoxide anions released by HeLa cancer cells. Graphical abstract Schematic of the microstructure of FePO4/C and FePO4/rGO-C nanofibers, a photograph of cancer cells (HeLa), and the electrochemical response towards O2-⢠of the sensor.
Assuntos
Materiais Biomiméticos/química , Compostos Férricos/química , Grafite/química , Nanofibras/química , Superóxidos/química , Superóxidos/metabolismo , Sobrevivência Celular , Condutividade Elétrica , Eletroquímica , Células HeLa , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Limite de Detecção , OxirreduçãoRESUMO
Reactive oxygen species (ROS)-driven chemodynamic therapy has emerged as a promising anti-tumor strategy. However, the insufficient hydrogen peroxide (H2O2) supply in tumor microenvironment results in a low Fenton reaction rate and subsequently poor ROS production and therapeutic efficacy. Herein, we report on a new nanocomposite MIL-53@ZIF-67/S loaded with doxorubicin and glucose oxidase, which is decomposed under the acidic tumor microenvironment to release Fe3+, Co3+, glucose oxidase, and doxorubicin. The released content leads to synergistic anti-tumor effect through the following manners: 1) doxorubicin is directly used for chemotherapy; 2) Fe3+and Co3+ result in glutathione depletion and Fenton reaction activation through Fe2+ and Co2+ generation to achieve chemodynamic therapy; 3) glucose oxidase continuously catalyzes glucose consumption to induce starvation of the cancer cells, and 4) at the same time the produced gluconic acid and H2O2 significantly promote Fenton reaction and further boost chemodynamic therapy. This work not only demonstrates the high anti-tumor effect of the new nanocomposite, but also provides an innovative strategy for the development of a multi-in-one nanoplatform for cancer therapy.
Assuntos
Cobalto , Doxorrubicina , Ferro , Estruturas Metalorgânicas , Nanocompostos , Nanocompostos/química , Cobalto/química , Cobalto/farmacologia , Estruturas Metalorgânicas/química , Estruturas Metalorgânicas/farmacologia , Estruturas Metalorgânicas/síntese química , Ferro/química , Humanos , Doxorrubicina/farmacologia , Doxorrubicina/química , Catálise , Animais , Camundongos , Peróxido de Hidrogênio/química , Glucose Oxidase/metabolismo , Glucose Oxidase/química , Microambiente Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral , Espécies Reativas de Oxigênio/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Antineoplásicos/farmacologia , Antineoplásicos/química , Propriedades de Superfície , Tamanho da Partícula , Ensaios de Seleção de Medicamentos AntitumoraisRESUMO
Pyroptosis-related long-noncoding RNAs (PRlncRNAs) play an important role in cancer progression. However, their role in lung squamous cell carcinoma (LUSC) is unclear. A risk model was constructed using the least absolute shrinkage and selection operator (LASSO) Cox regression analysis based on RNA sequencing data from The Cancer Genome Atlas database. The LUSC cohort was divided into high- and low-risk groups based on the median risk score. For the prognostic value of the model, the Kaplan-Meier analysis, log-rank test, and Cox regression analysis were performed. A nomogram was constructed to predict the prognosis of patients, using a risk score and clinical parameters such as age, sex, clinical stage, and tumor node metastasis classification (TNM) stage. Afterwards, six common algorithms were employed to assess the invasion of immune cells. The Gene Set Enrichment Analysis (GSEA) was conducted to identify differences between patients at high and low risk. Furthermore, the pRRophetic package was employed to forecast the half-maximal inhibitory doses of prevalent chemotherapeutic drugs, while the tumor immune dysfunction and exclusion score was computed to anticipate the response to immunotherapy. The expression levels of the seven PRlncRNAs were examined in both LUSC and normal lung epithelial cell lines using RT-qPCR. Proliferation, migration, and invasion assays were also carried out to investigate the role of MIR193BHG in LUSC cells. Patients in the low-risk group showed prolonged survival in the total cohort or subgroup analysis. The Cox regression analysis showed that the risk model could act as an independent prognostic factor for patients with LUSC. The results of GSEA analysis revealed that the high-risk group showed enrichment of cytokine pathways, Janus tyrosine kinase/signal transducer and activator of the transcription signalling pathway, and Toll-like receptor pathway. Conversely, the low-risk group showed enrichment of several gene repair pathways. Furthermore, the risk score was positively correlated with immune cell infiltration. Moreover, patients in the high-risk category showed reduced responsiveness to conventional chemotherapeutic medications and immunotherapy. The majority of the long noncoding RNAs in the risk model were confirmed to be overexpressed in LUSC cell lines compared to normal lung epithelial cell lines by in vitro tests. Further studies have shown that downregulating the expression of MIR193BHG may inhibit the growth, movement, and infiltration capabilities of LUSC cells, whereas increasing the expression of MIR193BHG could enhance these malignant tendencies. This study found that PRlncRNAs were linked to the prognosis of LUSC patients. The risk model, evaluated across various clinical parameters and treatment modalities, shows potential as a future reference for clinical applications.
Assuntos
Carcinoma de Células Escamosas , Neoplasias Pulmonares , Piroptose , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/terapia , Neoplasias Pulmonares/mortalidade , Masculino , Feminino , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/terapia , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/mortalidade , Prognóstico , Piroptose/genética , Imunoterapia , Pessoa de Meia-Idade , Nomogramas , Regulação Neoplásica da Expressão Gênica , Biomarcadores Tumorais/genética , Idoso , Linhagem Celular TumoralRESUMO
Chemodynamic therapy has become an emerging cancer treatment strategy, in which tumor cells are killed through toxic reactive oxygen species (ROS), especially hydroxyl radicals (ËOH) produced by the Fenton reaction. Nevertheless, low ROS generation efficiency and ROS depletion by cellular antioxidant systems are still the main obstacles in chemodynamic therapy. In the present work, we propose a dually enhanced chemodynamic therapy obtained by inhibiting ËOH consumption and promoting ËOH production based on the administration of bimetallic sulfide Co3-xCuxS4 nanoparticles functionalized by polyethylene glycol. These bimetallic nanoparticles display glutathione depleting and photothermal properties. The nanoparticles are gradually degraded in a tumor microenvironment, resulting in Co2+ and Cu2+ release. The released Co2+ triggers a Fenton-like reaction that turns endogenous hydrogen peroxide into highly toxic ËOH. In the cellular environment, Cu2+ ions are reduced to Cu+ by endogenous GSH, which decreases the intracellular antioxidant capacity and additionally up-regulates ËOH production via the Cu+-induced Fenton-like reaction. Moreover, under near-infrared light irradiation, the bimetallic nanoparticles display a photothermal conversion efficacy of 46.7%, which not only improves chemodynamic therapy via boosting a Fenton-like reaction but results in photothermal therapy through hyperthermia. Both in vitro cancer cell killing and in vivo tumor ablation experiments show that the bimetallic nanoparticles display outstanding therapeutic efficacy and negligible systemic toxicity, indicating their anticancer potential.
Assuntos
Hipertermia Induzida , Neoplasias , Antioxidantes , Cobre/farmacologia , Cobre/uso terapêutico , Glutationa/metabolismo , Peróxido de Hidrogênio/metabolismo , Neoplasias/tratamento farmacológico , Polietilenoglicóis , Espécies Reativas de Oxigênio , SulfetosRESUMO
Atherosclerosis (AS) is a chronic cardiovascular disease endangering human health and is one of the most common causes of myocardial infarction and stroke. Macrophage polarization plays a vital role in regulating plaque stability. As an important component of sunlight, ultraviolet B (UVB) has been proven to promote vitamin D and nitric oxide synthesis. This research used an AS model in ApoE-/- mice to study the effects of UVB on macrophage polarization and atherosclerotic plaque stability. In vitro, UVB irradiation increased arginase-I (Arg-I, M2 macrophage) and macrophage mannose receptor (CD206) expression, while the expression of inducible nitric oxide synthase (iNOS) (M1 macrophage) and CD86 was decreased. UVB promoted Akt phosphorylation in vitro. In vivo, UVB irradiation promoted the stabilization of atherosclerotic lesion plaques, while the phenotype of M2 macrophages increased. Our research provides new evidence for UVB in preventing and treating atherosclerosis.
Assuntos
Aterosclerose , Placa Aterosclerótica , Camundongos , Humanos , Animais , Ativação de Macrófagos , Aterosclerose/metabolismo , Macrófagos/patologia , Placa Aterosclerótica/patologia , FenótipoRESUMO
An ultrasensitive lateral flow immunoassay (LFIA) strip has been developed based on a multifunctional photothermal contrast Fe3O4@Au supraparticle (Fe3O4@Au SP) for quantitative Ochratoxin A (OTA) detection. The Fe3O4@Au SP composite not only shows a better photothermal effect over Fe3O4 and gold nanoparticles, but possesses magnetic property and excellent ability to directly adsorb protein (antibody). Under 808 nm irradiation, photothermal images of the test strips are recorded by a portable Infrared thermal camera. A quantitative analysis is easily achieved based on the thermal changes, which are proportional to the concentrations of analytes. Under an optimal condition, a wide linear detection range from 1 pg mLâ»1 to 1µgmLâ»1 and a limit of detection (LOD) at 0.12 pg mLâ»1 have been achieved with OTA as a model analytes. The practical application potential has also been validated by detection of OTA in spiked corn, peanut, and soybean extractives with overall recoveries ranging from 98.6% to 115% and coefficient of variations (CVs) between 6.06% and 12.73%. The photothermal LFIA renders a rapid, sensitive, and quantitative bio-/chemo-sensing platform with only a portable laser source and a thermal camera.
Assuntos
Ouro , Nanopartículas de Magnetita , Anticorpos , Imunoensaio , Limite de DetecçãoRESUMO
Reactive oxygen species (ROS) with strong oxidability have been considered as effective agents for antitumor therapy through oxidative damage to lipids, proteins, DNA and RNA. In this work, a multifunctional hollow cobaltosic sulfide (Co3S4)/photosensitizer indocyanine green (ICG) nanocomplex (Co3S4-ICG) has been synthesized by efficiently loading ICG into the hollow Co3S4 to realize synergistic antitumor therapy via chemodynamic therapy (CDT), photodynamic therapy (PDT) and photothermal therapy (PTT) under near-infrared (808 nm) laser irradiation. Co3S4 nanoparticles would be degraded in tumor acidic microenvironment into Co2+, which locally triggers a Fenton-like reaction to produce cytotoxic hydroxyl radicals (OH) for CDT. Co3S4-ICG could also produce singlet oxygen (1O2) through a multi-step photochemical process for PDT under 808 nm laser irradiation. The slow release of ICG in the tumor region was achieved due to hollow-structured Co3S4 working as nanocarriers, and which has been proved an effective approach for combined CDT/PDT. In addition, Co3S4-ICG showed high photothermal conversion efficiency (40.5%) for PTT, and excellent OH generation capability via photothermal-improved Fenton reaction, leading to the synergistically improved antitumor efficacy. In vitro and in vivo experimental results confirm that the combined PTT/PDT/photothermal-enhanced CDT therapy can effectively ablate tumors with a negligible systemic toxicity. This work provides a valuable strategy for designing and constructing of a multifunctional nanoplatform for synergistic antitumor therapy of solid tumors.