RESUMEN
Targeted protein degradation technologies (e.g., PROTACs) that can selectively degrade intracellular protein are an emerging class of promising therapeutic modalities. Herein, we describe the conjugation of photosensitizers and protein ligands (PS-Degrons), as an activable targeted protein degradation platform. PS-Degrons are capable of degrading protein of interest via light-triggered 1O2, which is orthogonal and complementary to existing technologies. This generalizable platform allows controllable knockdown of the target protein with high spatiotemporal precision. Our lead compound PSDalpha induces a complete degradation of human estrogen receptor α (ERα) under visible light. The high degrading ERα efficacy of PSDalpha enables an excellent anti-proliferation performance on MCF-7 cells. Our results establish a modular strategy for the controllable degradation of target proteins, which can hopefully overcome the systemic toxicity in clinical treatment of PROTACs. We anticipate that PS-Degrons would open a new chapter for biochemical research and for the therapeutics.
Asunto(s)
Proteínas/efectos de la radiación , Especies Reactivas de Oxígeno/química , Especies Reactivas de Oxígeno/efectos de la radiación , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Receptor alfa de Estrógeno/efectos de los fármacos , Receptor alfa de Estrógeno/metabolismo , Femenino , Humanos , Ligandos , Luz , Células MCF-7 , Modelos Moleculares , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/farmacología , Proteínas/química , Ubiquitina-Proteína LigasasRESUMEN
Light-emitting diode (LED)-based therapies, particularly blue LEDs with wavelengths of 400-500 nm, have shown beneficial results in several cancers, including melanoma, lymphoid cells, and skin tumors. In this study, the cell viability and apoptosis of Kasumi-1 cells treated by blue light (BL) irradiation have been explored. Firstly, BL can specially inhibit the proliferation and promote the apoptosis of Kasumi-1 cells. Furthermore, the apoptosis was triggered by the production of reactive oxygen species and the decline of mitochondrial membrane potential which was regulated by the ratio of Bcl-2(Bcl-xL)/Bax; BL caused the cells' final apoptosis accompanied with the increased cleavage of caspase-3 and poly-ADP-ribose polymerase. Finally, BL induced the degradation of AML1-ETO dependent on the activation of caspase-3. These results are helpful for establishing a low toxicity and high efficiency strategy of BL irradiation for clinical treatment of Kasumi-1 cells.
Asunto(s)
Apoptosis/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , Potencial de la Membrana Mitocondrial/efectos de la radiación , Proteínas de Fusión Oncogénica/metabolismo , Proteína 1 Compañera de Translocación de RUNX1/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Caspasa 3/metabolismo , Línea Celular Tumoral , Color , Subunidad alfa 2 del Factor de Unión al Sitio Principal/efectos de la radiación , Humanos , Proteínas de Fusión Oncogénica/efectos de la radiación , Estimulación Luminosa/métodos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Proteína 1 Compañera de Translocación de RUNX1/efectos de la radiaciónRESUMEN
BACKGROUND: Preclinical development of new drugs for cancer immunotherapy requires preconditioning total body irradiation (TBI) of mice to be humanized via hematopoietic stem cell transplantation. To assess the effect of preconditioning TBI, we detected the reactive oxygen species (ROS), Annexin V, propidium iodide (PI) level in bone marrow samples by flow cytometer. METHODS: We divided all NOG mice between irradiated (n = 20) and control groups (n = 10) for two time points. Irradiated mice were exposed to 3.5 Gy of radiation. After sacrificing BM samples were collected, the flow cytometric percentage of ROS, Annexin V, and PI markers were investigated on days 2 and 14 after exposure. RESULTS: At the first time point, the level of ROS was higher in the irradiated group than in the control group, and this difference was statistically significant (P < 0.05). Also, at the second time point, the mean differences of all markers in the irradiated group were significantly compared to the control group (P < 0.05). CONCLUSION: Thus, in NOG mice, the measurement of ROS level is helpful to the assessment of preconditioning TBI.
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Citometría de Flujo , Especies Reactivas de Oxígeno/efectos de la radiación , Irradiación Corporal Total/efectos adversos , Animales , Anexina A5/efectos de la radiación , Médula Ósea/efectos de la radiación , Trasplante de Células Madre Hematopoyéticas , Ratones , Propidio/efectos de la radiaciónRESUMEN
Radiation is considered as a promising insect pest control strategy for minimizing postharvest yield losses. Among various techniques, irradiation is a method of choice as it induces lethal biochemical or molecular changes that cause a downstream cascade of abrupt physiological abnormalities at the cellular level. In this study, we evaluated the effect of 60Co-γ radiation on various developmental stages of Zeugodacus cucurbitae Coquillett and subsequent carry-over effects on the progeny. For this purpose, we treated eggs with 30- and 50-Gy radiation doses of 60Co-γ. We found that radiation significantly affected cellular antioxidants, insect morphology, and gene expression profiles. Our results indicate that in response to various doses of irradiation reactive oxygen species, catalase, peroxidase, and superoxide dismutase activities were increased along with a significant increase in the malondialdehyde (MDA) content. We observed higher mortality rates during the pupal stage of the insects that hatched from irradiated eggs (50 Gy). Furthermore, the life span of the adults was reduced in response to 50 Gy radiation. The negative effects carried over to the next generation were marked by significantly lower fecundity in the F1 generation of the irradiation groups as compared to control. The radiation induced morphological abnormalities at the pupal, as well as the adult, stages. Furthermore, variations in the gene expression following irradiation are discussed. Taken together, our results signify the utility of 60Co-γ radiation for fruit fly postharvest management.
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Apoptosis/efectos de la radiación , Rayos gamma , Expresión Génica/efectos de la radiación , Tephritidae/efectos de la radiación , Animales , Antioxidantes/metabolismo , Antioxidantes/efectos de la radiación , Apoptosis/genética , Catalasa/metabolismo , Catalasa/efectos de la radiación , Radioisótopos de Cobalto/farmacología , Control de Insectos/métodos , Proteínas de Insectos/metabolismo , Proteínas de Insectos/efectos de la radiación , Larva/genética , Larva/metabolismo , Larva/fisiología , Larva/efectos de la radiación , Longevidad/efectos de la radiación , Malondialdehído/metabolismo , Malondialdehído/efectos de la radiación , Peroxidasa/metabolismo , Peroxidasa/efectos de la radiación , Control de Plagas/métodos , Pupa/genética , Pupa/metabolismo , Pupa/fisiología , Pupa/efectos de la radiación , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Tephritidae/genética , Tephritidae/metabolismo , Tephritidae/fisiologíaRESUMEN
Non-ionizing radiation is commonly used in the clinical setting, despite its known ability to trigger oxidative stress and apoptosis, which can lead to damage and cell death. Although induction of cell death is typically considered harmful, apoptosis can also be beneficial in the right context. For example, cell death can serve as the signal for new tissue growth, such as in apoptosis-induced proliferation. Recent data has shown that exposure to non-ionizing radiation (such as weak static magnetic fields, weak radiofrequency magnetic fields, and weak electromagnetic fields) is able to modulate proliferation, both in cell culture and in living organisms (for example during tissue regeneration). This occurs via in vivo changes in the levels of reactive oxygen species (ROS), which are canonical activators of apoptosis. This review will describe the literature that highlights the tantalizing possibility that non-ionizing radiation could be used to manipulate apoptosis-induced proliferation to either promote growth (for regenerative medicine) or inhibit it (for cancer therapies). However, as uncontrolled growth can lead to tumorigenesis, much more research into this exciting and developing area is needed in order to realize its promise.
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Apoptosis/efectos de la radiación , Proliferación Celular/efectos de la radiación , Radiación no Ionizante , Especies Reactivas de Oxígeno/efectos de la radiación , Animales , Protocolos Antineoplásicos , Humanos , Medicina RegenerativaRESUMEN
A new silicon phthalocyanine with piperazine-furan ring and its quaternized form were synthesized. All compounds were analyzed by spectroscopic techniques (FT-IR, 1H-NMR, MS, and UV-vis), and the absorbance characteristics of silicon phthalocyanines were evaluated with the expected strong typical absorption bands in the far-red spectrum. The cytotoxic effects of these phthalocyanines induced by photodynamic therapy (PDT) were determined in a dose-dependent manner. Following cytotoxicity analysis, flow cytometric research of cell death was performed. The formation of reactive oxygen species (ROS) was determined by confocal microscopy. High levels of cytotoxicity and decreased viable cell population have been detected in cancer cells after treatment. In addition, ROS formation was observed in PDT treated cancer cells. However, low levels of cell death and ROS formation were observed in non-tumorigenic cells. According to western blot data, PDT-mediated treatment was found to provide different expression patterns of the cleaved PARP1 protein. The presented study demonstrates that PDT-mediated treatment of newly synthesized phthalocyanines has significant anti-cancer effects on breast cancer cells and may induce different cell death pathways.
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Antineoplásicos/química , Neoplasias de la Mama/terapia , Indoles/química , Compuestos de Organosilicio/química , Fármacos Fotosensibilizantes/química , Piperazina/química , Apoptosis/efectos de la radiación , Línea Celular Tumoral , Regulación de la Expresión Génica/efectos de la radiación , Humanos , Indoles/farmacología , Imagen Óptica , Compuestos de Organosilicio/farmacología , Fotoquimioterapia , Fármacos Fotosensibilizantes/farmacología , Poli(ADP-Ribosa) Polimerasa-1/genética , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiaciónRESUMEN
BACKGROUND: Radiotherapy-induced tumor death remains critical in the successful first-line management of glioblastoma, whereas resistance to radiation serves as a major factor in disease progression. Mesenchymal shift has been identified as a driver in GBM recurrence, with gene expression associated with enhanced repair of macromolecular damage caused by radiation. METHODS: Using distinct mesenchymal subtype GBM cells lines, radiation response was assessed by clonogenic assay and orthotopic mouse tumor model. RNA-sequencing was performed in the setting of increasing radiation dosing while real-time assessment of ROS generation was achieved by the measurement of hydroxyl spin trap adducts via electron paramagnetic resonance. RESULTS: Radiation-induced cell death determined by clonogenic assay was significantly different at low dose (4-8 Gy) between the resistant U3035 cells and the sensitive U3020 cells. Similar trends were present in the in vivo NSG mouse model following radiation dosing on post-implantation day 7-10, with the rate of reduction in tumor bioluminescence reversing between the U3020 and U3035 cells after the third dose of radiation. Changes in gene expression following radiation determined by RNA-sequencing indicate both U3035 and U3020 cells demonstrate a shift toward more mesenchymal profiles, with concurrent shift away from pro-neural subtype gene expression in the U3020 cells that appeared to develop resistance to radiation in vivo. Persistence of ROS generated following radiation was greater in U3020 cells shown to be more sensitive to radiation. CONCLUSIONS: Despite the same molecular classification, distinct GBM cell lines can demonstrate differential response to radiation and potential for mesenchymal shift associated with radiation resistance.
Asunto(s)
Neoplasias Encefálicas/patología , Glioblastoma/patología , Tolerancia a Radiación/fisiología , Especies Reactivas de Oxígeno/efectos de la radiación , Transcriptoma/efectos de la radiación , Animales , Línea Celular Tumoral , Humanos , Ratones , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Glutamine metabolism provides energy to tumor cells and also produces reactive oxygen species (ROS). Excessive accumulation of ROS can damage mitochondria and eventually lead to cell death. xCT (SLC7A11) is responsible for the synthesis of glutathione in order to neutralize ROS. In addition, mitophagy can remove damaged mitochondria to keep the cell alive. Ionizing radiation kills tumor cells by causing the accumulation of ROS, which subsequently induces nuclear DNA damage. With this in mind, we explored the mechanism of intracellular ROS accumulation induced by ionizing radiation and hypothesized new methods to enhance the effect of radiotherapy. We used MCF-7 breast cancer cells and HCT116 colorectal cancer cells in our study. The above-mentioned cells were irradiated with different doses of X-rays or carbon ions. Clone formation assays were used to detect cell proliferation, enzyme-linked immunosorbent assay (ELISA) detected ATP, and glutathione (GSH) production, while the expression of proteins was detected by Western blot and quantitative real-time PCR analysis. The production of ROS was detected by flow cytometry, and immunofluorescence was used to track mitophagy-related processes. Finally, BALB/C tumor-bearing nude mice were irradiated with X-rays in order to further explore the protein expression found in tumors with the use of immunohistochemistry. Ionizing radiation increased the protein expressions of ASCT2, GLS, and GLUD in order to upregulate the glutamine metabolic flux in tumor cells. This caused an increase in ATP secretion. Meanwhile, ionizing radiation inhibited the expression of the xCT (SLC7A11) protein and reduced the generation of glutathione, leading to excessive accumulation of intracellular ROS. The mitophagy inhibitor, or knockdown Parkin gene, is able to enhance the ionizing radiation-induced ROS production and increase nucleus DNA damage. This combined treatment can significantly improve the killing effect of radiation on tumor cells. We concluded that ionizing radiation could upregulate the glutamine metabolic flux and enhance ROS accumulation in mitochondria. Ionizing radiation also decreased the SLC7A11 expression, resulting in reduced GSH generation. Therefore, inhibition of mitophagy can increase ionizing radiation-induced cell death.
Asunto(s)
Muerte Celular/efectos de la radiación , Glutamina/metabolismo , Glutamina/efectos de la radiación , Radiación Ionizante , Especies Reactivas de Oxígeno/efectos de la radiación , Animales , Femenino , Humanos , Ratones , Ratones Desnudos , Regulación hacia ArribaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Ultra Violet (UV) radiation is the major reason for reactive oxygen species (ROS) forming, skin cell damage, melanin production, and could horribly cause skin cancer. Saussureae Involucratae Herba (SIH) is the aerial part of Saussurea involucrata Matsum. & Koidz. This Material Medica is popular with both in Uyghur and Chinese medicines filed. SIH is one of the famous species of the Asteraceae family and which prescribed for skin protection from UV-induced damage according to China Pharmacopeia (2020). However, the detailed working mechanism involved is still not elucidated. AIM OF THE STUDY: We would like to probe the potential transduction pathway of SIH against UV-induced skin cell damages in cultured B16F10 cells. METHODS: Western blot, luciferase assay, laser confocal, RT-PCR and flow cytometer were employed here to verify the protective pharmaceutical value of SIH in cultured B16F10 cells after UV pre-treatment. RESULTS: Our result revealed that SIH attenuates ROS formation after UV-induced damage in B16F10 cells in a dose-dependent manner. Moreover, the transcriptional and translational anti-oxidative encoding genes were up-regulated under the presence of SIH. Further studies showed that SIH activated transcriptional activity of anti-oxidant response element (ARE). Moreover, we found that SIH dramatically stimulates PI3K/Akt phosphorylation in cultured B16F10 cells, this result was further verified by its specific inhibitors, LY294002 and Tocris. CONCLUSION: Our findings concluded that SIH protect melanoma cells from UV damages via activating PI3K/Akt signaling and which could provide scientific evidence for anti-UV pharmaceutical values of this herbal extract.
Asunto(s)
Fosfatidilinositol 3-Quinasas/metabolismo , Extractos Vegetales/farmacología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Saussurea , Transducción de Señal/efectos de los fármacos , Rayos Ultravioleta/efectos adversos , Animales , Melanoma Experimental , Ratones , Fosfatidilinositol 3-Quinasas/efectos de la radiación , Extractos Vegetales/uso terapéutico , Proteínas Proto-Oncogénicas c-akt/efectos de la radiación , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Transducción de Señal/fisiología , Transducción de Señal/efectos de la radiaciónRESUMEN
PURPOSE: To investigate whether the vascular collapse in tumors by conventional dose rate (CONV) irradiation (IR) would also occur by the ultra-high dose rate FLASH IR. METHODS AND MATERIALS: Lewis lung carcinoma (LLC) cells were subcutaneously implanted in mice. This was followed by CONV or FLASH IR at 15 Gy. Tumors were harvested at 6 or 48 hours after IR and stained for CD31, phosphorylated myosin light chain (p-MLC), γH2AX (a surrogate marker for DNA double strand break), intracellular reactive oxygen species (ROS), or immune cells such as myeloid and CD8α T cells. Cell lines were irradiated with CONV IR for Western blot analyses. ML-7 was intraperitoneally administered daily to LLC-bearing mice for 7 days before 15 Gy CONV IR. Tumors were similarly harvested and analyzed. RESULTS: By immunostaining, we observed that CONV IR at 6 hours resulted in constricted vessel morphology, increased expression of p-MLC, and much higher numbers of γH2AX-positive cells in tumors, which were not observed with FLASH IR. Mechanistically, MLC activation by ROS is unlikely, because FLASH IR produced significantly more ROS than CONV IR in tumors. In vitro studies demonstrated that ML-7, an inhibitor of MLC kinase, abrogated IR-induced γH2AX formation and disappearance kinetics. Lastly, we observed that CONV IR when combined with ML-7 produced some effects similar to FLASH IR, including reduction in the vasculature collapse, fewer γH2AX-positive cells, and increased immune cell influx to the tumors. CONCLUSIONS: FLASH IR produced novel changes in the tumor microenvironment that were not observed with CONV IR. We believe that MLC activation in tumors may be responsible for some of the microenvironmental changes differentially regulated between CONV and FLASH IR.
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Carcinoma Pulmonar de Lewis/radioterapia , Cadenas Ligeras de Miosina/efectos de la radiación , Microambiente Tumoral/efectos de la radiación , Animales , Azepinas/administración & dosificación , Vasos Sanguíneos/patología , Vasos Sanguíneos/efectos de la radiación , Linfocitos T CD8-positivos/citología , Carcinoma Pulmonar de Lewis/irrigación sanguínea , Carcinoma Pulmonar de Lewis/metabolismo , Histonas/metabolismo , Histonas/efectos de la radiación , Masculino , Ratones , Ratones Endogámicos C57BL , Cadenas Ligeras de Miosina/antagonistas & inhibidores , Cadenas Ligeras de Miosina/metabolismo , Naftalenos/administración & dosificación , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/efectos de la radiación , Radioterapia/métodos , Dosificación Radioterapéutica , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiaciónRESUMEN
Low level light therapy uses light of specific wavelengths in red and near-infrared spectral range to treat various pathological conditions. This light is able to modulate biochemical cascade reactions in cells that can have important health implications. In this study, the effect of low intensity light at 650, 808 and 1064 nm on neurons and two types of cancer cells (neuroblastoma and HeLa) is reported, with focus on the photoinduced change of intracellular level of Ca2+ ions and corresponding signaling pathways. The obtained results show that 650 and 808 nm light promotes intracellular Ca2+ elevation regardless of cell type, but with different dynamics due to the specificities of Ca2+ regulation in neurons and cancer cells. Two origins responsible for Ca2+ elevation are determined to be: influx of exogenous Ca2+ ions into cells and Ca2+ release from endoplasmic reticulum. Our investigation of the related cellular processes shows that light-induced membrane depolarization is distinctly involved in the mechanism of Ca2+ influx. Ca2+ release from endoplasmic reticulum activated by reactive oxygen species generation is considered as a possible light-dependent signaling pathway. In contrast to the irradiation with 650 and 808 nm light, no effects are observed under 1064 nm irradiation. We believe that the obtained insights are of high significance and can be useful for the development of drug-free phototherapy.
Asunto(s)
Señalización del Calcio/efectos de la radiación , Calcio/efectos de la radiación , Retículo Endoplásmico/metabolismo , Retículo Endoplásmico/efectos de la radiación , Calcio/fisiología , Membrana Celular/metabolismo , Electrofisiología , Colorantes Fluorescentes/química , Células HeLa , Humanos , Rayos Infrarrojos , Terapia por Luz de Baja Intensidad , Neuronas/efectos de la radiación , Imagen Óptica , Especies Reactivas de Oxígeno/efectos de la radiaciónRESUMEN
Differential inherent and acquired radioresistance of human lung cancer cells contribute to poor therapeutic outcome and tumor recurrence after radiotherapy. Inherent radioresistance of lung cancer cells is known to be associated with ROSLow cancer stem cells (CSCs). However, mechanism of acquired radioresistance in lung cancer cells is poorly understood. Here, we exposed human lung cancer cells (A549) to a cumulative dose of 40Gy and allowed the radioresistant (RR) survivors to divide and form macroscopic colonies after each fraction of 5Gy dose. The RR subline exhibited enrichment of cytosolic ROSHigh cells without specific increase in mitochondrial ROS levels. We found a concomitant increase in the expression of redox regulatory transcription factor Nrf2 and its dependent antioxidant genes in RR cells and cell cycle delay as compared to parental cells. The treatment of RR cells with Nrf2 inhibitor resulted in decreased clonogenic survival indicating their addiction to Nrf2 for metabolic adaptations under high levels of cytosolic ROS. A causal role of inherent ROS levels in conferring radioresistance was established by sorting ROSHigh and ROSLow populations from parental and RR cells. It was observed that ROSHigh population from both parental and RR cells exhibited radioresistance as observed by clonogenic assay. Interestingly, ROSHigh population of cells exhibited higher levels of cellular thiols in both parental and RR cells. Thus, our observations highlight presence of a novel subpopulation in lung cancer cells, which exhibits radioresistance by maintaining 'oxidative stress' and Nrf2 dependent metabolic adaptations. We also posit Nrf2 pathway as a druggable target for radiosensitization of RR A549 cells.
Asunto(s)
Adaptación Fisiológica/efectos de la radiación , Radioisótopos de Cobalto/química , Resistencia a Antineoplásicos/efectos de la radiación , Neoplasias Pulmonares/radioterapia , Estrés Oxidativo/efectos de la radiación , Células A549 , Antioxidantes/metabolismo , Apoptosis/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Regulación de la Expresión Génica/efectos de la radiación , Humanos , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , Células Madre Neoplásicas/efectos de la radiación , Oxidación-Reducción , Tolerancia a Radiación , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Compuestos de Sulfhidrilo/metabolismoRESUMEN
Here, we report the synthesis of a quantum dot (QD)-DNA covalent conjugate to be used as an H2O2-free DNAzyme system with oxidase activity. Amino-coupling conjugation was carried out between amino-modified oligonucleotides (CatG4-NH2) and carboxylated quantum dots (CdTe@COOH QDs). The obtained products were characterized by spectroscopic methods (UV-Vis, fluorescence, circular dichroizm (CD), and IR) and the transmission electron microscopy (TEM) technique. A QD-DNA system with a low polydispersity and high stability in aqueous solutions was successfully obtained. The catalytic activity of the QD-DNA conjugate was examined with Amplex Red and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)) indicators using reactive oxygen species (ROS) generated by visible light irradiation. The synthesized QD-DNAzyme exhibited enhanced catalytic activity compared with the reference system (a mixture of QDs and DNAzyme). This proved the assumption that the covalent attachment of DNAzyme to the surface of QD resulted in a beneficial effect on its catalytic activity. The results proved that the QD-DNAzyme system can be used for generation of the signal by light irradiation. The light-induced oxidase activity of the conjugate was demonstrated, proving that the QD-DNAzyme system can be useful for the development of new cellular bioassays, e.g., for the determination of oxygen radical scavengers.
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ADN Catalítico/metabolismo , Oxidorreductasas/metabolismo , Puntos Cuánticos , Benzotiazoles/química , Benzotiazoles/metabolismo , ADN Catalítico/química , Luz , Microscopía Electrónica de Transmisión , Nanopartículas/química , Nanopartículas/ultraestructura , Oxazinas/química , Oxazinas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Análisis Espectral/métodos , Ácidos Sulfónicos/química , Ácidos Sulfónicos/metabolismoRESUMEN
A continuous state of oxidative stress during inflammation contributes to the development of 25% of human cancers. Epithelial and inflammatory cells release reactive oxygen species (ROS) and reactive nitrogen species (RNS) that can damage DNA. ROS/RNS have biological implications in both chemoresistance and tumor recurrence. As several clinically employed anticancer drugs can generate ROS/RNS, we have addressed herein how inducible nitric oxide synthase and nitric oxide (iNOS/â¢NO) affect the molecular pathways implicated in the tumor response to oxidative stress. To mimic the oxidative stress associated with chemotherapy, we used a photosensitizer (pheophorbide a) that can generate ROS/RNS in a controlled manner. We investigated how iNOS/â¢NO modulates the tumor response to oxidative stress by involving the NF-κB and Nrf2 molecular pathways. We found that low levels of iNOS induce the development of a more aggressive tumor population, leading to survival, recurrence and resistance. By contrast, high levels of iNOS/â¢NO sensitize tumor cells to oxidative treatment, causing cell growth arrest. Our analysis showed that NF-κB and Nrf2, which are activated in response to oxidative stress, communicate with each other through RKIP. For this critical role, RKIP could be an interesting target for anticancer drugs. Our study provides insight into the complex signaling response of cancer cells to oxidative treatments as well as new possibilities for the rational design of new therapeutic strategies.
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Óxido Nítrico/fisiología , Estrés Oxidativo/fisiología , Neoplasias de la Próstata/metabolismo , Fármacos Sensibilizantes a Radiaciones/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Línea Celular Tumoral , Clorofila/análogos & derivados , Clorofila/toxicidad , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Humanos , Masculino , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico/efectos de la radiación , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico Sintasa de Tipo II/efectos de la radiación , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/efectos de la radiación , Neoplasias de la Próstata/patología , Especies Reactivas de Oxígeno/efectos de la radiaciónRESUMEN
Ionizing radiation exposure produces direct or indirect biological effects on genomic DNA. The latter are ionizing radiation mediated by induction of free radicals and oxygen species (ROS). The study was conducted to evaluate the dose-effect/time-effect of antioxidant treatments in reducing the induction of double-strand breaks in human blood lymphocytes. Human peripheral blood samples of 2 mL each from healthy donors were irradiated with 10 mGy after pre-incubation with different antioxidants (ß-carotene, vitamin E, vitamin C, N-acetyl L-cysteine). In order to assess their efficiency as prophylactic therapy for irradiation, various concentrations and combinations of antioxidants, as well as different incubation times, have been evaluated. To assess double-strand breaks induced by ionizing radiation, the phosphorylated histone γ-H2AX has been used. A significant reduction (p < 0.001) in double-strand breaks studied with a γ-H2AX assay was observed with N-acetyl L-cysteine with a 1-h incubation time, followed by vitamin C, vitamin E, and ß-carotene. The use of antioxidants, especially N-acetyl L-cysteine before irradiation, significantly decreased the occurrence of double-strand breaks, demonstrating the potential radiological protection for exposure to ionizing radiation.
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Antioxidantes/farmacología , Células Sanguíneas/efectos de la radiación , Roturas del ADN de Doble Cadena/efectos de la radiación , ADN/efectos de la radiación , Histonas/efectos de la radiación , Linfocitos/efectos de la radiación , Acetilcisteína/farmacología , Ácido Ascórbico/farmacología , Relación Dosis-Respuesta en la Radiación , Histonas/genética , Humanos , Microscopía Fluorescente , Dosis de Radiación , Radiación Ionizante , Radiografía , Especies Reactivas de Oxígeno/efectos de la radiación , Vitamina E/farmacología , Rayos X , beta Caroteno/farmacologíaRESUMEN
BACKGROUND: Antioxidase alleviates the accumulation of radiation-induced reactive oxygen species (ROS) and therefore has strong connections with radioresistance. Isocitrate dehydrogenase 2 (IDH2) facilitates the turnover of antioxidase, but its role in radiotherapeutic efficiency in oesophageal squamous cell carcinoma (ESCC) still remains elusive. METHODS: The involvement of IDH2 in radiotherapeutic efficacy in ESCC was investigated in vitro and vivo by IDH2 knockdown. IDH2 expression in biopsy specimens of 141 patients was identified to evaluate its clinical significance. RESULTS: We found that Kyse510 and Kyse140 cells were more radioresistant and had higher IDH2 expression. In these two cell lines, IDH2 knockdown intensified the radiation-induced ROS overload and oxidative damage on lipid, protein, and nucleic acids. In addition, IDH2 silencing aggravated the radiation-induced mitochondrial dysfunction and cell apoptosis and ultimately promoted radiosensitisation via inhibiting AKT phosphorylation in a ROS-dependent manner. Furthermore, IDH2 depletion facilitated the radiation-induced growth inhibition and cell apoptosis in murine xenografts. Finally, IDH2 expression was correlated with definite chemoradiotherapy (dCRT) efficacy and served as an independent prognostic factor for survival of ESCC patients. CONCLUSIONS: IDH2 plays a key role in the radioresistance of ESCC. Targeting IDH2 could be a promising regimen to improve radiotherapeutic efficiency in ESCC patients.
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Carcinoma de Células Escamosas de Esófago/radioterapia , Isocitrato Deshidrogenasa/genética , Estrés Oxidativo/efectos de la radiación , Tolerancia a Radiación/genética , Animales , Antioxidantes/metabolismo , Apoptosis/efectos de la radiación , Carcinoma de Células Escamosas de Esófago/genética , Carcinoma de Células Escamosas de Esófago/patología , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Xenoinjertos , Humanos , Masculino , Ratones , Ratones Noqueados , Mitocondrias/genética , Mitocondrias/efectos de la radiación , Estrés Oxidativo/genética , Especies Reactivas de Oxígeno/efectos de la radiaciónRESUMEN
UVA irradiation induced ROS-mediated photo damage to the human skin leading to coarseness, wrinkling, pigmentation, and cutaneous malignancies. We investigated the dermatoprotective efficacies of submicromolar concentrations of ergothioneine (EGT, 0.125-0.5 µM), which occurs naturally as a sulfur-containing amino acid, in the mechanisms in human skin fibroblast (HSF) cells. UVA-induced AP-1 (c-Fos and c-Jun) translocation was found to be inhibited by EGT treatments with the parallel inhibition of the collagenolytic matrix metalloproteinase- (MMP-) 1 activation and type I procollagen degradation. Moreover, EGT mitigated UVA-induced ROS generation. An increase in the amount of antioxidant genes (HO-1, NQO-1, and γ-GCLC) from EGT and were associated with upregulated Nrf2 expressions in a dose-dependent or time-dependent manner. We confirmed this from Nrf2 translocation and increased nuclear ARE promoter activity that underlie EGT dermatoprotective activities. Also, glutathione (GSH) levels (from γ-GCLC) were significantly increased. Moreover, we showed that mediated by ERK, JNK, and PKC, signaling cascades mediate Nrf2 translocation. We confirmed this phenomenon by the suppressed nuclear Nrf2 activation in cells that were treated with respective inhibitors (PD98059, SP600125, and GF109203X). However, antioxidant protein expressions were impaired in Nrf2 knockdown cells to confirm that ARE/Nrf2 pathways and the inhibition of AP-1 had significant roles in EGT-mediated protective effects. We can conclude that ergothioneine ameliorated UVA-induced skin aging and is a useful food supplement for skin care products.
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Ergotioneína/farmacología , Fibroblastos/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Transducción de Señal/efectos de los fármacos , Envejecimiento de la Piel/efectos de los fármacos , Factor de Transcripción AP-1/metabolismo , Antioxidantes/farmacología , Línea Celular , Fibroblastos/citología , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Glutatión/metabolismo , Hemo-Oxigenasa 1/metabolismo , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Factor 2 Relacionado con NF-E2/genética , Procolágeno/metabolismo , Proteína Quinasa C/metabolismo , ARN Interferente Pequeño , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Transducción de Señal/genética , Factor de Transcripción AP-1/antagonistas & inhibidores , Rayos Ultravioleta , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Previous studies have shown that curcumin (Cur) induced by ultrasound has protective effects on atherosclerosis even if low bioavailability of the Cur. The enhancement of bioavailability of the Cur further improved the curative effect of sonodynamic therapy (SDT) on atherosclerosis through nanotechnology. Nanosuspensions as a good drug delivery system had obvious advantages in increasing the solubility and improving the effectiveness of insoluble drugs. The aim of this study was to develop curcumin nanosuspensions (Cur-ns) which used polyvinylpyrrolidone (PVPK30) and sodium dodecyl sulfate (SDS) as stabilizers to improve poor water solubility and bioavailability of the Cur. And then the therapeutic effects of Cur-ns-SDT on atherosclerotic plaques and its possible mechanisms would be investigated and elucidated. Cur-ns with a small particle size has been successfully prepared and the data have confirmed that Cur-ns could be more easily engulfed into RAW264.7 cells than free Cur and accumulated more under the stimulation of the ultrasound. Reactive oxygen species (ROS) inside RAW264.7 cells after SDT led to the decrease of mitochondrial membrane potential (MMP) and the higher expression of cleaved caspase-9/3. The results of in vivo experiments showed that Cur-ns-SDT reduced the level of total cholesterol (TC) and low density lipoprotein (LDL) and promoted the transformation from M1 to M2 macrophages, relieved atherosclerosis syndrome. Therefore, Cur-ns-SDT was a potential treatment of anti-atherosclerosis by enhancing macrophages apoptosis through mitochondrial pathway and inhibiting the progression of plaques by interfering with macrophages polarization.
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Aterosclerosis/terapia , Curcumina/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Nanomedicina Teranóstica/métodos , Terapia por Ultrasonido/métodos , Administración Oral , Animales , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Aterosclerosis/sangre , Disponibilidad Biológica , Colesterol/sangre , Terapia Combinada/métodos , Curcumina/farmacocinética , Modelos Animales de Enfermedad , Humanos , Lipoproteínas LDL/sangre , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Potencial de la Membrana Mitocondrial/efectos de la radiación , Ratones , Ratones Noqueados para ApoE , Nanopartículas/química , Tamaño de la Partícula , Vehículos Farmacéuticos/química , Povidona/química , Células RAW 264.7 , Especies Reactivas de Oxígeno/metabolismo , Especies Reactivas de Oxígeno/efectos de la radiación , Dodecil Sulfato de Sodio/químicaRESUMEN
Low Temperature Plasma (LTP) generates reactive oxygen and nitrogen species, causing cell death, similarly to radiation. Radiation resistance results in tumour recurrence, however mechanisms of LTP resistance are unknown. LTP was applied to patient-derived prostate epithelial cells and gene expression assessed. A typical global oxidative response (AP-1 and Nrf2 signalling) was induced, whereas Notch signalling was activated exclusively in progenitor cells. Notch inhibition induced expression of prostatic acid phosphatase (PAP), a marker of prostate epithelial cell differentiation, whilst reducing colony forming ability and preventing tumour formation. Therefore, if LTP is to be progressed as a novel treatment for prostate cancer, combination treatments should be considered in the context of cellular heterogeneity and existence of cell type-specific resistance mechanisms.
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Gases em Plasma/uso terapéutico , Neoplasias de la Próstata/radioterapia , Tolerancia a Radiación/efectos de la radiación , Receptores Notch/genética , Fosfatasa Ácida/genética , Muerte Celular/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Línea Celular Tumoral , Proliferación Celular/efectos de la radiación , Células Epiteliales/efectos de la radiación , Regulación Neoplásica de la Expresión Génica/efectos de la radiación , Humanos , Masculino , Factor 2 Relacionado con NF-E2/genética , Gases em Plasma/efectos adversos , Próstata/patología , Próstata/efectos de la radiación , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Tolerancia a Radiación/genética , Especies de Nitrógeno Reactivo/efectos de la radiación , Especies Reactivas de Oxígeno/efectos de la radiación , Transducción de Señal/efectos de la radiación , Células Madre/efectos de la radiación , Factor de Transcripción AP-1/genéticaRESUMEN
Literature studies suggest important protective effects of low-frequency, low-energy pulsed electromagnetic fields (PEMFs) on inflammatory pathways affecting joint and cerebral diseases. However, it is not clear on which bases they affect neuroprotection and the mechanism responsible is yet unknown. Therefore the aim of this study was to identify the molecular targets of PEMFs anti-neuroinflammatory action. The effects of PEMF exposure in cytokine production by lipopolysaccharide (LPS)-activated N9 microglial cells as well as the pathways involved, including adenylyl cyclase (AC), phospholipase C (PLC), protein kinase C epsilon (PKC-ε) and delta (PKC-δ), p38, ERK1/2, JNK1/2 mitogen activated protein kinases (MAPK), Akt and caspase 1, were investigated. In addition, the ability of PEMFs to modulate ROS generation, cell invasion and phagocytosis, was addressed. PEMFs reduced the LPS-increased production of TNF-α and IL-1ß in N9 cells, through a pathway involving JNK1/2. Furthermore, they decreased the LPS-induced release of IL-6, by a mechanism not dependent on AC, PLC, PKC-ε, PKC-δ, p38, ERK1/2, JNK1/2, Akt and caspase 1. Importantly, a significant effect of PEMFs in the reduction of crucial cell functions specific of microglia like ROS generation, cell invasion and phagocytosis was found. PEMFs inhibit neuroinflammation in N9 cells through a mechanism involving, at least in part, the activation of JNK MAPK signalling pathway and may be relevant to treat a variety of diseases characterized by neuroinflammation.