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1.
Anticancer Res ; 40(1): 229-238, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31892571

RESUMO

BACKGROUND/AIM: We previously reported the potential of aminonaphthoquinone derivatives as therapeutic agents against breast and other oestrogen-responsive tumours when combined with curcumin. This study aimed at screening of novel aminonaphthoquinone derivatives (Rau 008, Rau 010, Rau 015 and Rau 018) combined with curcumin for cytotoxic, anti-angiogenic and anti-metastatic effects on MCF-7 and MDA-MB-231 breast cancer cells. MATERIALS AND METHODS: Cytotoxic and anti-angiogenic effects were analysed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and enzyme-linked immunosorbent assay; while anti-metastatic effects were measured using adhesion assay, Boyden chambers and Matrigel. RESULTS: Curcumin combined with Rau 008 elicited marked cytotoxic effects in MCF-7 cells compared with the individual treatments, whereas when it was combined with Rau 015 and with Rau 018, it displayed similar effects in MDA-MB-231 cells. The anti-angiogenic effect of Rau 015 plus curcumin in MCF-7 cells and Rau 018 plus curcumin in MDA-MB-231 cells was more effective than individual treatments, while the metastatic capability of MDA-MB-231 cells was significantly reduced after treatment with the aminonaphthoquinone-curcumin combinations. CONCLUSION: Aminonaphthoquinones may offer significant promise as therapeutic agents against breast cancer, particularly when combined with curcumin.


Assuntos
Neoplasias da Mama/patologia , Curcumina/farmacologia , Progressão da Doença , Naftoquinonas/farmacologia , Neoplasias da Mama/irrigação sanguínea , Neoplasias da Mama/tratamento farmacológico , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Curcumina/química , Curcumina/uso terapêutico , Matriz Extracelular/metabolismo , Feminino , Humanos , Células MCF-7 , Naftoquinonas/química , Invasividade Neoplásica , Neovascularização Patológica/tratamento farmacológico , Fator A de Crescimento do Endotélio Vascular/metabolismo
2.
Toxicol Lett ; 322: 120-130, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31953210

RESUMO

Strong epidemiological evidence supports the association between increased air pollution and the risk of developing atherosclerotic cardiovascular diseases (CVDs). However, the mechanism remains unclear. As an environmental air pollutant and benzo-a-pyrene (BP) metabolite, BP-1,6-quinone (BP-1,6-Q) is present in the particulate phase of air pollution. This study was undertaken to examine the redox activity of BP-1,6-Q and mechanisms associated with it using EA.hy926 endothelial cells. BP-1,6-Q at 0.01-1 µM significantly stimulated the production of reactive oxygen species (ROS)·in intact cells and isolated mitochondria. Furthermore, BP-1,6-Q-induced ROS was altered by mitochondrial electron transport chain (METC) inhibitors of complex I (rotenone) and complex III (antimycin A), denoting the involvement of mitochondrial electron transport chain (METC) in BP-1,6-Q mediated ROS production. In METC deficient cells, interestingly, BP-1,6-Q-mediated ROS production was enhanced, suggesting that overproduction of ROS by BP-1,6-Q is not only produced from mitochondria but can also be from the cell outside of mitochondria (extramitochondrial). BP-1,6-Q also triggered endothelial-monocyte interaction and stimulated expression of vascular adhesion molecule-1 (VCAM-1). In conclusion, these results demonstrate that BP-1,6-Q can generate ROS within both mitochondria and outside of mitochondria, resulting in stimulation of adhesion of monocytes to endothelial cells, a key event in the pathogenesis of atherosclerosis.


Assuntos
Benzopirenos/toxicidade , Células Endoteliais/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Técnicas de Cocultura , Complexo de Proteínas da Cadeia de Transporte de Elétrons/metabolismo , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Monócitos/metabolismo , Oxirredução , Transdução de Sinais , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/metabolismo
3.
J Photochem Photobiol B ; 202: 111718, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31790883

RESUMO

Metallic nanoparticles were extensively examined to explore their impending exploitations over pharmaceutical purposes. Current work attempting to explores the cytotoxic capacity of zinc oxide (ZnO) nanoparticles besides to human melanoma cell line (A375). Viability of cells was resoluted, and the promising cytotoxicity potential was exhibited by zinc oxide nanoparticles. Cellular adhesion and morphology was determined by propidium iodide assay. Characterization studies like UV-Spectroscopy, X-ray diffraction (XRD) investigation, transmission electron microscope (TEM), energy dispersive X-ray (EDX) Spec, and Fourier transform infrared (FT-IR) examination confirms the accessibility of measurement, form and volume. The mRNA expression of apoptotic genes like caspase 3, 8 and 9 was elevated followed by the exposure to ZnO nanoparticles and it was narrowly proved that ZnO nanoparticles stimulates the apoptotic cell necrosis at the transcriptional stage. Cardiospermum halicacabum down regulated the apoptotic gene expressions. Reactive oxygen species (ROS) accumulation was augmented at concentration reliant mode, that changed normalize numerous indicator pathways and manipulate the kinetic cellular actions. ZnO nanoparticle synthesized Cardiospermum halicacabum might persuades programmed cell necrosis via elevated ROS levels in cells. CH-ZnONPs was further stimulates the markers of apoptosis and aggravates necrosis of cancerous cells, toxicity to cells, and accretion of ROS. With sourced on above whole data, this might accomplished that CH-ZnONPs amalgamated Cardiospermum halicacabum appreciably possessed a toxicity to human melanoma cells (A375) via provoking the apoptotic cell necrosis, entailed feasible efficacy of CH-ZnONPs besides malignancy management.


Assuntos
Antineoplásicos/síntese química , Apoptose , Nanopartículas Metálicas/química , Sapindaceae/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Química Verde , Humanos , Melanoma/metabolismo , Melanoma/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Extratos Vegetais/química , Folhas de Planta/química , Folhas de Planta/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sapindaceae/metabolismo , Óxido de Zinco/química
4.
J Photochem Photobiol B ; 202: 111644, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31770706

RESUMO

Management of degenerative spine pathologies frequently leads to the need for bone growth. Rehmanniae Radix (RR), a Chinese herbal formulation was found to exhibit numerous therapeutic properties including its potent effect against cancer cell lines. However, the underlying mechanism through which the Zinc oxide nanoparticles (ZnONPs) synthesized from Rehmanniae Radix exerts its anti-cancer activity against osteosarcoma cell line MG-63 needs to be explored. Therefore, the study was performed to evaluate the anticancer, cytotoxicity and apoptotic effectiveness of ZnONPs from RR against MG-63 cells. Characterization studies such UV-vis spectroscopy, FTIR, TEM and XRD analysis were performed. Cytotoxicity assay, mitochondrial membrane potential (MMP), morphological examination of cells and formation of reactive oxygen species (ROS), and apoptosis inducing ability of RR were evaluated by various procedures. Western blot analysis of apoptotic markers such as Bax, caspase-3 and caspase-9 were also performed. RR was found to inhibit growth of MG-63 cells at increasing dose. AO/EB staining confirmed the apoptotic efficacy of ZnONPs induced by RR in MG-63 cells. ZnONPs was also found to initiate increased generation of ROS and decreased MMP. Decreased MMP has resulted in increased levels of apoptotic proteins Bax, caspase-3 and caspase-9 and induction of apoptosis was substantiated by western blot analysis. The outcomes of the work propose that ZnONPs from RR exhibits strong anticancer action and inducing apoptosis on MG-63 cells via stimulating increased generation of ROS. Thus, ZnONPs from RR might be used as a hopeful drug target against several types of cancer cell lines.


Assuntos
Caspase 3/metabolismo , Química Verde , Nanopartículas Metálicas/química , Óxido de Zinco/química , Proteína X Associada a bcl-2/metabolismo , Caspase 3/genética , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Extratos Vegetais/química , Espécies Reativas de Oxigênio/metabolismo , Rehmannia/química , Rehmannia/metabolismo , Proteína X Associada a bcl-2/genética
5.
J Photochem Photobiol B ; 202: 111677, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31810037

RESUMO

In this examination, chitosan-silk fibroin/polyethylene terephthalate (CTS-SF/PET), chitosan-silk fibroin/polyethylene terephthalate/hydroxyapatite (CTS-SF/PET/HAP) and chitosan-silk fibroin/polyethylene terephthalate/Silver @hydroxyapatite (CTS-SF/PET/Ag@HAP) scaffolds were prepared by utilizing the plasma splashing procedure. Field emission scanning electron microscopy (FESEM) results demonstrated that the outside of the PET covered with HAP nanoparticles. The cell viability results demonstrated that the number of Mesenchymal stem cells (MSCs) primarily spread out on CTS-SF/PET/Ag@HAP. RT-PCR results demonstrated that there was an upregulated mRNA articulation of osseous development-related properties in the CTS-SF/PET/Ag@HAP composite. The in vivo rabbit animal assessment scores of the CTS-SF/PET/Ag@HAP composite were significantly better than those of the CTS-SF/PET at 1 to 3 months. Both in-vivo and in-vitro results exhibited in this investigation recommend that the cytocompatibility and osseointegration of CTS-SF/PET/Ag@HAP tendon were fundamentally improved by expanding the multiplication of cells and up-regulating the outflow of tendon development-related properties. In conclusion, the CTS-SF/PET/Ag@HAP tendon is a promising candidate for Anterior Cruciate Ligament (ACL) replacement in the future.


Assuntos
Ligamento Cruzado Anterior/fisiologia , Materiais Biocompatíveis/química , Durapatita/química , Nanopartículas/química , Osseointegração , Prata/química , Animais , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Artropatias/reabilitação , Artropatias/terapia , Artropatias/veterinária , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Nanopartículas/toxicidade , Osseointegração/efeitos dos fármacos , Coelhos , Tecidos Suporte/química
6.
Gut ; 69(2): 252-263, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31092589

RESUMO

OBJECTIVE: To study the role of α4ß7 integrin for gut homing of monocytes and to explore the biological consequences of therapeutic α4ß7 inhibition with regard to intestinal wound healing. DESIGN: We studied the expression of homing markers on monocyte subsets in the peripheral blood and on macrophage subsets in the gut of patients with IBD and controls with flow cytometry and immunohistochemistry. Integrin function was addressed with dynamic adhesion assays and in vivo gut homing assays. In vivo wound healing was studied in mice deficient for or depleted of α4ß7 integrin. RESULTS: Classical and non-classical monocytes were clearly dichotomous regarding homing marker expression including relevant expression of α4ß7 integrin on human and mouse non-classical monocytes but not on classical monocytes. Monocyte-expressed α4ß7 integrin was functionally important for dynamic adhesion to mucosal vascular addressin cell adhesion molecule 1 and in vivo gut homing. Impaired α4ß7-dependent gut homing was associated with reduced (effect size about 20%) and delayed wound healing and suppressed perilesional presence of wound healing macrophages. Non-classical monocytes in the peripheral blood were increased in patients with IBD under clinical treatment with vedolizumab. CONCLUSION: In addition to reported effects on lymphocytes, anti-α4ß7 therapy in IBD also targets non-classical monocytes. Impaired gut homing of such monocytes might lead to a reduction of wound healing macrophages and could potentially explain increased rates of postoperative complications in vedolizumab-treated patients, which have been observed in some studies.


Assuntos
Doenças Inflamatórias Intestinais/patologia , Integrinas/fisiologia , Intestinos/patologia , Monócitos/fisiologia , Cicatrização/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Monoclonais Humanizados/farmacologia , Estudos de Casos e Controles , Adesão Celular/efeitos dos fármacos , Adesão Celular/fisiologia , Quimiotaxia de Leucócito/fisiologia , Feminino , Fármacos Gastrointestinais/farmacologia , Humanos , Doenças Inflamatórias Intestinais/sangue , Doenças Inflamatórias Intestinais/fisiopatologia , Integrinas/antagonistas & inibidores , Integrinas/sangue , Mucosa Intestinal/metabolismo , Intestinos/fisiologia , Masculino , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Monócitos/metabolismo , Cicatrização/efeitos dos fármacos , Adulto Jovem
7.
Biosci Biotechnol Biochem ; 84(1): 103-110, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31559912

RESUMO

We previously reported that MDA-MB-231 and MCF-7 cells, which are breast cancer cell lines and have cancer and cancer-initiating cells (CICs), were killed following normothermic microwave irradiation in which the cellular temperature was maintained at 37°C. In this study, we investigated the percentages of live or dead cells among CD44+/CD24- cells, which were defined as CICs among MDA-MB-231 and MCF-7 cells, and other types of cells in response to microwave irradiation. CD44+/CD24- cells among MDA-MB-231 cells were killed, thereby decreasing the number of cells, whereas the number of live CD44+/CD24- MCF-7 cells was increased following microwave irradiation. Moreover, adhesion, invasion, and migration were decreased in MDA-MB-231 cells, and the activation of matrix metalloproteinase-2 (MMP-2) in MDA-MB-231 cells was increased following microwave irradiation. These decreased cell activities might have been caused by MMP-2 activation and population changes in CD44+/CD24- in MDA-MB-231 cells.Abbreviations: APC: allophecocyanin; CBB: coomassie Brilliant Blue; CD: cluster of differentiation; CICs: cancer-initiating cells; FACS: fluorescence-activated cell sorting; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; FTDT: finite-difference time domain; HER2: human epidermal growth factor receptor type 2; PI: propidium iodide.


Assuntos
Antígeno CD24/metabolismo , Adesão Celular/efeitos da radiação , Movimento Celular/efeitos da radiação , Receptores de Hialuronatos/metabolismo , Micro-Ondas , Neoplasias de Mama Triplo Negativas/patologia , Apoptose/efeitos da radiação , Contagem de Células , Corantes/metabolismo , Feminino , Citometria de Fluxo , Humanos , Células MCF-7 , Metaloproteinase 2 da Matriz/metabolismo , Invasividade Neoplásica , Propídio/metabolismo , Temperatura Ambiente
8.
J Photochem Photobiol B ; 203: 111744, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31887637

RESUMO

Articular cartilage regeneration is a challenging process due to its inadequate ability of self-recovering biological mechanisms. The progresses of cartilage tissue engineering is supported to overwhelmed the repairing difficulties and degenerative diseases. The main goal of the present study is to design biomaterials with suitable physico-chemical, mechanical and biological properties for the carrier of growth factor and improving differentiation of mesenchymal stem cell into damaged cartilage tissues. Herein, TGF-ß loaded hydrogel network was prepared through the chemical interactions between vinyl group of natural polymers. Fourier-transform infrared spectroscopy results show the characteristic peaks at 3074 cm-1, 1713 cm-1, and 810 cm-1, which confirm the existence of the vinyl group and successful formation of maleoyl functionalized Chitosan (MCh). The obtained MCh was freely dissolved in the distilled water up to 8% (w/v). X-ray photoelectron spectroscopy survey spectral results show a peak at 289.0 eV which revealed that the OCO and DS were 1.2% and also evidenced the methacryl substitution of Silk fibroin (SF) nanoformulations. The weight loss and mechanical test were analyzed and the results showed that MSF acts as a foremost crosslinking point with MCh through the reaction between the methacrylate groups of MSF and maleoyl groups of MCh which led to enhancing the density and improved the compressive strength. The maximum drug release activity was recorded in the TGF-ß loaded MCh@MSF hydrogel compared to bare MCh hydrogel. Further, the TGF-ß loaded MCh@ MSF hydrogel exhibited the cell viability percentage nearly at 79-102% for MC3T3-E1 and 88-104% for BMDSCs. Similarly, the TGF-ß loaded MCh@MSF exhibited the highest inhibitory activity against E. coli (83%) than S. aureus (67%). Overall, this study concluded the TGF-ß loaded MCh@MSF showed better biocompatibility and could be utilized in the field of cartilage tissue engineering.


Assuntos
Cartilagem Articular/fisiologia , Quitosana/química , Hidrogéis/química , Regeneração , Seda/química , Fator de Crescimento Transformador beta/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Hidrogéis/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Nanopartículas/química , Regeneração/efeitos dos fármacos , Reologia , Staphylococcus aureus/efeitos dos fármacos , Engenharia Tecidual , Fator de Crescimento Transformador beta/química
9.
Immunology ; 159(1): 63-74, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31573680

RESUMO

Monocyte-derived macrophages (MDMs) generated from peripheral blood monocytes are widely used to model human macrophages for in vitro studies. However, the possible impact of different isolation methods on the resulting MDM phenotype is poorly described. We aimed to investigate the effects of three commonly used monocyte isolation techniques on the resulting MDM phenotype. Plastic adhesion, negative selection, and CD14pos selection were compared. Monocyte-derived macrophages were generated by 5-day culture with macrophage and granulocyte-macrophage colony-stimulating factors. We investigated monocyte and MDM yields, purity, viability, and cell phenotype. CD14pos selection resulted in highest monocyte yield (19·8 × 106 cells, equivalent to 70% of total) and purity (98·7%), compared with negative selection (17·7 × 106 cells, 61% of total, 85·0% purity), and plastic adhesion (6·1 × 106 cells, 12·9% of total, 44·2% purity). Negatively selected monocytes were highly contaminated with platelets. Expression of CD163 and CD14 were significantly lower on CD14pos selection and plastic adhesion monocytes, compared with untouched peripheral blood mononuclear cells. After maturation, CD14pos selection also resulted in the highest MDM purity (98·2%) compared with negative selection (94·5%) and plastic adhesion (66·1%). Furthermore, MDMs from plastic adhesion were M1-skewed (CD80high  HLA-DRhigh  CD163low ), whereas negative selection MDMs were M2-skewed (CD80low  HLA-DRlow  CD163high ). Choice of monocyte isolation method not only significantly affects yield and purity, but also impacts resulting phenotype of cultured MDMs. These differences may partly be explained by the presence of contaminating cells when using plastic adherence or negative selection. Careful considerations of monocyte isolation methods are important for designing in vitro assays on MDMs.


Assuntos
Diferenciação Celular , Separação Celular/métodos , Citometria de Fluxo , Receptores de Lipopolissacarídeos/metabolismo , Macrófagos/fisiologia , Monócitos/fisiologia , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Biomarcadores , Adesão Celular , Células Cultivadas , Humanos , Interleucina-6/metabolismo , Lectinas Tipo C/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Lectinas de Ligação a Manose/metabolismo , Monócitos/imunologia , Monócitos/metabolismo , Fenótipo , Receptores de Superfície Celular/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
10.
J Colloid Interface Sci ; 559: 65-75, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31610306

RESUMO

Electroactive nanofibrous scaffold is a vital tool for the study of the various biological research fields from bioelectronics to regenerative medicine, which can provide cell preferable 3D nanofiber architecture and programmed electrical signal. However, intrinsic non-biodegradability is a major problem that hinders its widespread application in the clinic. Herein, we designed, synthesized, and characterized shell/core poly (3,4-ethylenedioxythiophene) (PEDOT)/chitosan (CS) nanofibers by combining the electrospinning and recrystallization processes. Upon incorporating a trace amount of PEDOT (1.0 wt%), the resultant PEDOT/CS nanofibers exhibited low interfacial charge transfer impedance, high electrochemical stability, high electrical conductivity (up to 0.1945 S/cm), and ultrasensitive piezoelectric property (output voltage of 22.5 mV by a human hair prodding). With such unique electrical and conductive properties, PEDOT/CS nanofibers were further applied to brain neuroglioma cells (BNCs) to stimulate their adhesion, proliferation, growth, and development under an optimal external electrical stimulation (ES) and a pulse voltage of 400 mV/cm. ES-responsive PEDOT/CS nanofibers indeed promoted BNCs growth and development as indicated by a large number and density of axons. The synergetic interplay between external ES and piezoelectric voltage demonstrates new PEDOT-based nanofibers as implantable electroactive scaffolds for numerous applications in nerve tissue engineering, human health monitoring, brain mantle information extraction, and degradable microelectronic devices.


Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/química , Quitosana/química , Condutividade Elétrica , Nanofibras/química , Polímeros/química , Testes de Impedância Acústica/métodos , Axônios/metabolismo , Materiais Biocompatíveis/química , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cristalização , Estimulação Elétrica/métodos , Glioma/metabolismo , Humanos
11.
Biosci Biotechnol Biochem ; 84(1): 85-94, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31794329

RESUMO

Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) regulates collagen-mediated platelet activation through its cytoplasmic immunoreceptor tyrosine-based inhibition motifs (ITIMs). However, the function of CEACAM1's extracellular cleavage fragments is currently unknown. In the present study, we used mass spectrometry (MS) to identify 9 cleavage fragments shed by matrix metallopeptidase 12 (MMP-12), and then we synthesized peptides with sequences corresponding to the fragments. QLSNGNRTLT (QLSN), a peptide from the A1-domain of CEACAM1, significantly attenuated collagen-induced platelet aggregation. QLSN also attenuated platelet static adhesion to collagen. Additionally, QLSN reduced human platelet secretion and integrin αIIbß3 activation in response to glycoprotein VI (GPVI)-selective agonist, convulxin. Correspondingly, QLSN treatment significantly decreased convulxin-mediated phosphorylation of Src, protein kinase B (Akt), spleen tyrosine kinase (Syk) and phospholipase Cγ2 (PLCγ2) in human platelets. These data indicate that the CEACAM1-derived peptide QLSN inhibits GPVI-mediated human platelet activation. QLSN could potentially be developed as a novel antiplatelet agent.


Assuntos
Antígenos CD/metabolismo , Moléculas de Adesão Celular/metabolismo , Colágeno/metabolismo , Oligopeptídeos/farmacologia , Ativação Plaquetária/efeitos dos fármacos , Glicoproteínas da Membrana de Plaquetas/metabolismo , Plaquetas/metabolismo , Adesão Celular/efeitos dos fármacos , Venenos de Crotalídeos/farmacologia , Humanos , Motivo de Inibição do Imunorreceptor Baseado em Tirosina/fisiologia , Lectinas Tipo C , Metaloproteinase 12 da Matriz/metabolismo , Oligopeptídeos/síntese química , Fosfolipase C gama/metabolismo , Fosforilação/efeitos dos fármacos , Agregação Plaquetária/efeitos dos fármacos , Glicoproteínas da Membrana de Plaquetas/agonistas , Domínios Proteicos/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quinase Syk/metabolismo
12.
Bioelectrochemistry ; 132: 107435, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31855831

RESUMO

Cell electrotransfection is an effective approach for transferring exogenous molecules into living cells by electric stimulation. The existing in-situ electrotransfection micro-devices for adherent cells exhibit the drawbacks of low transfection efficiency and low cell viability. An important reason for these drawbacks is the unequal exposure of cells to the electric field. It was found that cells growing directly below the energized electrodes experience a much lower electric field intensity when compared to the cells growing below the spacing area of the electrodes, resulting in low transfection with a strip-like pattern. Therefore, a new strategy for the in-situ electrotransfection of adherent cells growing in a standard 12-well plate is proposed in this study. By sequentially energizing electrodes arranged in a nested and non-contact manner, the cells were exposed to an overall equal intensity of the electric field, and thus a higher efficiency of transfection was achieved. The seven cell lines transfected using this method exhibited high transfection efficiency and high cell viability, demonstrating the potential for studying gene function.


Assuntos
Adesão Celular , Eletrodos , Eletroporação/métodos , Transfecção/métodos , Sobrevivência Celular , Células HeLa , Humanos
13.
Talanta ; 207: 120259, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31594574

RESUMO

We report a new method: biomimetic cell-cell adhesion capillary electrophoresis (BCCACE) to screen drugs targeting interactions between cell membrane receptors and ligands under an environment close to physiological conditions, in which the cell membrane receptors/ligands can maintain their natural conformations and bioactivity without being isolated and purified. Firstly, we screened twenty-one lactose derivatives by cell-immobilized capillary electrophoresis and obtained Gu-4 with the best activity (K = 3.58 ±â€¯0.22 × 104) targeting macrophage antigen-1 (Mac-1). Then, BCCACE was performed as follows: HEK 293 cells overexpressed with receptor (intercellular adhesion molecules-1, ICAM-1) were cultured and immobilized on the inner wall of capillaries as stationary phase, which simulated the endothelial cells lining on the inner surface of blood vessels. HEK 293 cells overexpressed with ligand Mac-1 as samples were used to simulate the neutrophils cells in blood vessels. And Gu-4 added into the running buffer solution as the antagonist was used to simulate the drug in blood. The results showed that Gu-4 (40 µM) could selectively inhibit cell-cell adhesion by targeting the interaction between Mac-1 and ICAM-1. Finally, the pharmaceutical efficacy assays of Gu-4 at cellular and animal levels were carried out using the concentration of 40 µM and the dose of 20 mg kg-1 respectively, which showed the anti-cancer metastasis activity of Gu-4 and the validity of the method. This method simulated a complete three-dimensional vascular model, which can easily obtain the suitable blood concentration of drugs. This system simulated the interaction between leukocytes and vascular endothelial cells in the bloodstream antagonized by drugs, and obtained the effective concentration of the antagonist. It can be used as an accuracy and efficient drug screening method and will be expected to become a new method to screen drugs targeting cell-cell adhesion.


Assuntos
Biomimética/métodos , Adesão Celular/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Eletroforese Capilar/métodos , Glutamina/análogos & derivados , Lactose/análogos & derivados , Proteínas de Membrana/metabolismo , Relação Dose-Resposta a Droga , Glutamina/farmacologia , Células HEK293 , Humanos , Lactose/farmacologia , Ligantes , Ligação Proteica/efeitos dos fármacos , Cicatrização/efeitos dos fármacos
14.
Eur J Med Chem ; 185: 111823, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31698158

RESUMO

Breast cancer is the most frequently diagnosed malignancy and the second common cause of death in women worldwide. High mortality in breast cancer is frequently associated with metastatic progression rather than the primary tumor itself. It has been recently identified that the CXCR4/CXCL12 axis plays a pivotal role in breast cancer metastasis, especially in directing metastatic cancer cells to CXCL12-riched organs and tissues. Herein, taking the amide-sulfamide as the lead structure, the second-round structural modifications to the sulfamide structure were performed to obtain more active CXCR4 modulators against tumor metastasis. Both in vivo and in vitro experiments illustrated that compound IIIe possessed potent CXCR4 binding affinity, excellent anti-metastatic and anti-angiogenetic activity against breast cancer. More importantly, in a mouse breast cancer lung metastasis model, compound IIIe exerted a significant inhibitory effect on breast cancer metastasis. Taken together, all these positive results demonstrated that developing of CXCR4 modulators is a promising strategy to mediate breast cancer metastasis.


Assuntos
Amidas/farmacologia , Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Quimiocina CXCL12/antagonistas & inibidores , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Experimentais/tratamento farmacológico , Receptores CXCR4/antagonistas & inibidores , Amidas/administração & dosagem , Amidas/química , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Neoplasias da Mama/patologia , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quimiocina CXCL12/metabolismo , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Humanos , Injeções Intravenosas , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Nus , Estrutura Molecular , Neoplasias Experimentais/patologia , Neoplasias Experimentais/secundário , Receptores CXCR4/metabolismo , Relação Estrutura-Atividade , Células Tumorais Cultivadas , Cicatrização/efeitos dos fármacos
15.
Anticancer Res ; 39(12): 6915-6921, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31810962

RESUMO

BACKGROUND/AIM: The present study aimed to investigate the role of Homebox B2 (HOXB2) in bladder cancer (BC). MATERIALS AND METHODS: The Cancer Genome Atlas (TCGA) dataset was used to analyse HOXB2 expression in BC. The influence of HOXB2 on the cellular functions of BC cells was determined in both HOXB2 knockdown and HOXB2 overexpressed BC cell lines using in vitro assays. RESULTS: HOXB2 mRNA was significantly upregulated in luminal infiltrated and luminal papillary subtypes of BC. Drug Metabolism Cytochrome P450 was significantly enriched in BCs expressing high levels of HOXB2. Knockdown of HOXB2 from EJ138 cells reduced growth, adhesion and invasion. In contrast, overexpression of HOXB2 in RT112 cells induced growth and adhesion of bladder cancer cells. CONCLUSION: Increased HOXB2 expression in papillary BC can promote cell growth and adhesion of BC cells. Drug Metabolism Cytochrome P450 pathway was enriched in BCs overexpressing HOXB2.


Assuntos
Carcinoma Papilar/genética , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Regulação para Cima , Neoplasias da Bexiga Urinária/genética , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sistema Enzimático do Citocromo P-450/genética , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Invasividade Neoplásica
16.
Phys Rev Lett ; 123(22): 228102, 2019 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-31868410

RESUMO

Biological function requires cell-cell adhesions to tune their cohesiveness; for instance, during the opening of new fluid-filled cavities under hydraulic pressure. To understand the physical mechanisms supporting this adaptability, we develop a stochastic model for the hydraulic fracture of adhesive interfaces bridged by molecular bonds. We find that surface tension strongly enhances the stability of these interfaces by controlling flaw sensitivity, lifetime, and optimal architecture in terms of bond clustering. We also show that bond mobility embrittles adhesions and changes the mechanism of decohesion. Our study provides a mechanistic background to understand the biological regulation of cell-cell cohesion and fracture.


Assuntos
Adesão Celular/fisiologia , Junções Intercelulares/química , Junções Intercelulares/fisiologia , Modelos Biológicos , Membrana Celular/química , Membrana Celular/fisiologia , Simulação por Computador , Processos Estocásticos , Tensão Superficial
17.
Anticancer Res ; 39(11): 6015-6023, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704827

RESUMO

BACKGROUND/AIM: We previously reported that expression of melanoma-associated antigen (MAGE)-D4 mRNA was a prognostic factor for esophageal squamous cell carcinoma (ESCC). The aim of this study was to validate the expression of MAGE-D4 in two additional patient cohorts, and to investigate its biological functions. MATERIALS AND METHODS: The role of MAGE-D4 in cell proliferation, adhesion, and migration was determined by gene knockdown experiments in the KYSE590 cell line. MAGE-D4 protein expression was analyzed in ESCC tissues by immunohistochemistry. A second validation cohort consisted of an ESCC mRNA dataset from The Cancer Genome Atlas. RESULTS: Knockdown of MAGE-D4 significantly decreased cell proliferation and migration. Expression of MAGE-D4 protein was significantly associated with disease-free survival. In the second validation cohort, high MAGE-D4 mRNA expression was associated with significantly shorter overall survival and disease-free survival. CONCLUSION: MAGE-D4 plays an important role in the malignant behavior of ESCC. MAGE-D4 was validated as a prognostic indicator in two independent ESCC patient cohorts.


Assuntos
Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/secundário , Proteínas de Neoplasias/metabolismo , Idoso , Antígenos de Neoplasias/genética , Apoptose , Biomarcadores Tumorais/genética , Adesão Celular , Proliferação de Células , Estudos de Coortes , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/cirurgia , Carcinoma de Células Escamosas do Esôfago/metabolismo , Carcinoma de Células Escamosas do Esôfago/cirurgia , Feminino , Seguimentos , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Invasividade Neoplásica , Metástase Neoplásica , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Prognóstico , RNA Interferente Pequeno/genética , Taxa de Sobrevida , Células Tumorais Cultivadas
18.
Anticancer Res ; 39(11): 6135-6144, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704841

RESUMO

AIM: To investigate the effect of carnosine, an active compound of dietary beef, fish and chicken, on the regulation of cell adhesion and extravasation during metastasis. MATERIALS AND METHODS: Cell adhesion and extravasation abilities, and related regulating molecular mechanisms were analyzed in human colorectal cancer cells (HCT-116) and human umbilical vein cells (EA.hy926). RESULTS: Carnosine reduced the ability of HCT-116 cells to adhere to EA.hy926 cells. The expression levels of integrin-ß1 in HCT-116 cells, as well as of intercellular adhesion molecule-1 and E-selectin in EA.hy926 cells, were reduced after carnosine treatment. After EA.hy926 cells were treated with carnosine, phosphorylation of vascular endothelia-cadherin (VE-cadherin), protein levels of Ras homologous (RHO) and RHO-associated coiled-coil containing protein kinase, and levels of reactive oxygen species were reduced. After treating EA.hy926 cells with carnosine, phosphorylation of inhibitor of kappa B (IκB) and DNA binding activity of nuclear factor-κB (NF-κB) were reduced. CONCLUSION: Carnosine inhibits metastatic cell adhesion and extravasation by suppressing NF-κB signaling activation.


Assuntos
Carnosina/farmacologia , Adesão Celular/efeitos dos fármacos , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/metabolismo , Humanos , NF-kappa B/genética , NF-kappa B/metabolismo , Células Tumorais Cultivadas
19.
Int J Nanomedicine ; 14: 6615-6630, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31695360

RESUMO

Background: Nanocomposites produced by reinforcement of polysaccharide matrix with nanoparticles are widely used in engineering of biomaterials. However, clinical applications of developed novel biomaterials are often limited due to their poor biocompatibility. Purpose: The aim of this work was to comprehensively assess biocompatibility of highly macroporous chitosan/agarose/nanohydroxyapatite bone scaffolds produced by a novel method combining freeze-drying with a foaming agent. Within these studies, blood plasma protein adsorption, osteoblast (MC3T3-E1 Subclone 4 and hFOB 1.19) adhesion and proliferation, and osteogenic differentiation of mesenchymal stem cells derived from bone marrow and adipose tissue were determined. The obtained results were also correlated with materials' surface chemistry and wettability to explain the observed protein and cellular response. Results: Obtained results clearly showed that the developed nanocomposite scaffolds were characterized by high biocompatibility and osteoconductivity. Importantly, the scaffolds also revealed osteoinductive properties since they have the ability to induce osteogenic differentiation (Runx2 synthesis) in undifferentiated mesenchymal stem cells. The surface of biomaterials is extremely hydrophilic, prone to protein adsorption with the highest affinity toward fibronectin binding, which allows for good osteoblast adhesion, spreading, and proliferation. Conclusion: Produced by a novel method, macroporous nanocomposite biomaterials have great potential to be used in regenerative medicine for acceleration of the bone healing process.


Assuntos
Regeneração Óssea , Osso e Ossos/fisiologia , Quitosana/química , Durapatita/química , Nanocompostos/química , Osteoblastos/citologia , Sefarose/química , Engenharia Tecidual/métodos , Tecidos Suporte/química , Adsorção , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Biomarcadores/metabolismo , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Cães , Feminino , Humanos , Células-Tronco Mesenquimais/citologia , Camundongos , Osteogênese/efeitos dos fármacos , Molhabilidade
20.
J Photochem Photobiol B ; 201: 111651, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31683166

RESUMO

Designing biomimetic biomaterials influenced by the common complex structure of hard tissues is yet a test these days. The control of bio-mineralization procedure onto biomaterials should be assessed before the use in medical applications. Coming to the bone rejuvenation applications, this work assessed the in vitro antibacterial activity and interacting between osteoblast cells (MG63) on poly (hydroxypropyl methacrylate) (PHPMA) cryogel consolidated with Zn/Ce substituted hydroxyapatite (MHAp) nanocomposite (PHPMA/MHAp). Osteoblast cell multiplication, morphology, and metabolic action were assessed through various conventions. The functional group, texture, mechanical properties, and protein adsorption profiles of the fabricated nanocomposite were analyzed by the FTIR, XRD, SEM, and mechanical examinations, respectively. The bacterial activity of nanocomposites was additionally assessed against E. coli and S. aureus microorganisms, individually. Nanocomposite advanced endo-chondral ossification at the messed up parts of the bone deformity than cryogel did. These results recommend that PHPMA/MHAp nanocomposites joined the good innate properties of each polymer and bioceramic, giving a mechanically powerful, cell-responsive, and permeable stage for hard tissue applications.


Assuntos
Criogéis/química , Durapatita/química , Fraturas do Fêmur/terapia , Nanocompostos/química , Nanopartículas/química , Animais , Regeneração Óssea/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Cério/química , Modelos Animais de Doenças , Escherichia coli/efeitos dos fármacos , Fraturas do Fêmur/patologia , Nanocompostos/uso terapêutico , Nanocompostos/toxicidade , Polímeros/química , Ratos , Staphylococcus aureus/efeitos dos fármacos , Engenharia Tecidual , Zinco/química
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