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1.
Nan Fang Yi Ke Da Xue Xue Bao ; 39(9): 1030-1037, 2019 Sep 30.
Artigo em Chinês | MEDLINE | ID: mdl-31640957

RESUMO

OBJECTIVE: To investigate the relationship between necroptosis and apoptosis in MCET3-E1 cell death induced by glucocorticoids. METHODS: MC3T3-E1 cells were incubated with 10-6 mol/L dexamethasone followed by treatment with the apoptosis inhibitor z-VAD-fmk (40 µmol/L) or the necroptosis inhibitor necrostatin-1 (40 µmol/L) for 2 h. At 72 h after incubation with dexamethasone, the cells were harvested to determine the cell viability using WST-1 assay and the rate of necrotic cells using annexin V/PI double staining; the percentage of apoptotic cells was determined using Hoechst staining. The mitochondrial membrane potential and the level of ATP in the cells were also evaluated. Transmission electron microscopy was used to observe the microstructural changes of the cells. The expressions of RIP-1 and RIP-3 in the cells were detected by Western blotting. RESULTS: At a concentration of 10-6 mol/L, dexamethasone induced both apoptosis and necroptosis in MC3T3- E1 cells. Annexin V/PI double staining showed that inhibition of cell apoptosis caused an increase in cell necrosis manifested by such changes as mitochondrial swelling and plasma membrane disruption, as shown by electron microscopy; Hoechst staining showed that the percentage of apoptotic cells was significantly reduced. When necroptosis was inhibited by necrostatin-1, MC3T3-E1 cells showed significantly increased apoptosis as shown by both AV/PI and Hoechst staining, and such changes were accompanied by changes in mitochondrial membrane potential and ATP level in the cells. CONCLUSIONS: In the process of dexamethasone-induced cell death, necroptosis and apoptosis can transform reciprocally accompanied by functional changes of the mitochondria.


Assuntos
Apoptose , Morte Celular/efeitos dos fármacos , Dexametasona , Necrose , Células 3T3 , Trifosfato de Adenosina , Animais , Potencial da Membrana Mitocondrial , Camundongos , Microscopia Eletrônica , Mitocôndrias/ultraestrutura
2.
J Agric Food Chem ; 67(37): 10321-10329, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31419115

RESUMO

Pterostilbene (PTS) is a phenolic compound with diverse pharmacologic activities. However, its potential for inhibiting obesity-related colorectal cancer (CRC) remains unclear. Our study evaluated the mechanism of inhibitory effects of PTS on adipocyte conditioned-medium (aCM)-induced malignant transformation in HT-29 colorectal adenocarcinoma cells. The results demonstrated that PTS could downregulate the expression of aCM-induced fatty acid-binding protein 5 (FABP5) and prometastatic factors such as vascular endothelial growth factor, matrix metalloproteinase-2 (MMP2), MMP9, and extracellular tumor necrosis factor α via inhibiting aCM-induced nuclear factor-kappa B (NF-κB), ß-catenin, and peroxisome proliferator-activated receptor γ (PPAR-γ). Moreover, PTS can suppress aCM-stimulated phosphoinositide 3-kinase (PI3K), protein kinase B (Akt), p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinases 1/2 (JNK 1/2) signaling pathways activation that are upstream of NF-κB, ß-catenin, and PPAR-γ. Therefore, we suggest that PTS could alleviate adiposity-induced metastasis in CRC via inhibiting cell migration through downregulating FABP5 gene expression.


Assuntos
Adipócitos/metabolismo , Movimento Celular/efeitos dos fármacos , Neoplasias Colorretais/fisiopatologia , Meios de Cultivo Condicionados/química , Proteínas de Ligação a Ácido Graxo/metabolismo , Estilbenos/farmacologia , Células 3T3 , Animais , Linhagem Celular , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Meios de Cultivo Condicionados/metabolismo , Regulação para Baixo/efeitos dos fármacos , Proteínas de Ligação a Ácido Graxo/genética , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , NF-kappa B/genética , NF-kappa B/metabolismo , PPAR gama/genética , PPAR gama/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
3.
Cell Biochem Funct ; 37(6): 400-407, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31328801

RESUMO

There are two types of bisphosphonates (BPs), nitrogen-containing (N-BPs) and those free from nitrogen (non-N-BPs). Although N-BPs show greater inhibition of bone resorption than non-N-BPs, their effects are likely accompanied with inflammation, which non-N-BPs mitigate. We examined the competitive effects of zoledronate (ZOL), an N-BP, and etidronate (ETI), a non-N-BP, in osteoblasts. ZOL, but not ETI, markedly reduced alkaline phosphatase activity and cell viability in osteoblastic MC3T3-E1 and Saos2 cells, while that inhibition was relieved by simultaneous administration of ETI, possibly because of competition with ZOL for cellular uptake. However, phosphonoformate, an inhibitor of the phosphonate transporters SLC20A and SLC34A, did not mitigate the reducing effects of ZOL, suggesting that those transporters are not involved in BP uptake in osteoblastic cells. Additionally, ZOL reduced fibroblastic NIH3T3 and C3H10T1/2 cell viability, which was relieved by administration of both ETI and phosphonoformate. Transporter gene expression levels were significantly lower in osteoblasts as compared with fibroblasts, which may account for the distinct effects of phosphonoformate with different cell types. Together, our results suggest existence of a common uptake route of N-BPs and non-N-BPs into osteoblastic cells that is unrelated to the SLC20A and SLC34A families. SIGNIFICANCE OF THE STUDY: N-BP ZOL was shown to suppress differentiation and viability of osteoblasts. ZOL-induced cell viability suppression was also observed in fibroblasts, which was markedly relieved by addition of the non-N-BP ETI. Additionally, mitigation of the effects of ZOL was achieved with phosphonoformate, a sodium-phosphate cotransporter inhibitor, in fibroblastic cells but not osteoblasts. Expression levels of SLC20A and SLC34A family genes were significantly lower in osteoblasts as compared with fibroblasts. These observations suggest that incorporation of N-BPs and non-N-BPs in osteoblasts is mediated via common transporters that appear to be distinct from SLC20A and 34A, which operate in fibroblasts.


Assuntos
Difosfonatos/farmacologia , Osteoblastos/efeitos dos fármacos , Proteínas Cotransportadoras de Sódio-Fosfato/antagonistas & inibidores , Células 3T3 , Animais , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Humanos , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Relação Estrutura-Atividade
4.
Biomater Sci ; 7(9): 3918-3925, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31322162

RESUMO

Polymer gels can be classified as chemical and physical gels, depending on the type of cross-link. Physical gels usually form by physical cross-linking, such as hydrogen bonding, van der Waals and/or p-p interactions, which can avoid the use of additional cross-linking agents. Polyamidoamine (PAMAM) dendrimers possess abundant active groups on their surfaces. Modified dendrimers have been used as versatile linkers in some projects. In this work, polymer gels composed of PAMAM dendrimers without any covalent bonding cross-linking agents were prepared. The number of amino groups and ester groups on the surface of the dendrimers was precisely regulated to help form hydrogen bonds between adjacent dendrimers. The prepared dendrimer-based polymer gels retain the properties of PAMAM dendrimers such as antibacterial properties, and the unique structures make the gels exhibit high compressive strengths but relatively low tensile strengths. Interestingly, the prepared gels show good anti-inflammatory properties in acute inflammation models of mice with ear edema. The inflammatory inhibition rate and hematoxylin-eosin (H&E) staining method were used to confirm the anti-inflammatory effect. This present study demonstrates that the dendrimer-based polymer gels achieved through hydrogen bonding could be a versatile platform for tissue engineering.


Assuntos
Anti-Inflamatórios/farmacologia , Dendrímeros/química , Poliaminas/química , Células 3T3 , Animais , Sobrevivência Celular/efeitos dos fármacos , Força Compressiva , Dendrímeros/farmacologia , Edema/tratamento farmacológico , Feminino , Géis , Ligações de Hidrogênio , Camundongos , Camundongos Endogâmicos BALB C , Poliaminas/farmacologia , Resistência à Tração
5.
Carbohydr Polym ; 219: 87-94, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31151549

RESUMO

Oxidized cellulose is the most used hemostatic materials in clinical applications. In addition to its perfect hemostatic efficiency, it is degradable under in vivo conditions and supremely prevents bacterial growth. On the other hand, one of the drawbacks of the oxidized cellulose is cytotoxicity due to the strongly acidic nature during degradation. There is a number of commercially available oxidized cellulose products which are derived from regenerated and non-regenerated cellulose. On the other hand, the effect of oxidation degree and structure (regenerated or non-regenerated) on product efficiency is undetermined. Moreover, oxidation degree which is primary factor for both bactericidal and hemostatic efficiency is also crucial for assessment of the product. In this study, oxidized cellulose versus oxidized regenerated cellulose microparticles with various oxidation degree was produced and characterized. Comparative studies were conducted in terms of bactericidal and hemostatic efficiencies in addition to cytotoxicity. The results could be a reference for the optimized oxidized cellulose product for the hemostatic applications.


Assuntos
Antibacterianos , Celulose Oxidada/farmacologia , Celulose Oxidada/toxicidade , Hemólise , Hemostasia/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Células 3T3 , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Celulose Oxidada/química , Humanos , Camundongos , Oxirredução
6.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 36(3): 421-427, 2019 Jun 25.
Artigo em Chinês | MEDLINE | ID: mdl-31232545

RESUMO

In this study, we aim to investigat the effect of microgravity on osteoblast differentiation in osteoblast-like cells (MC3T3-E1). In addition, we explored the response mechanism of nuclear factor-kappa B (NF-κB) signaling pathway to "zero- g" in MC3T3-E1 cells under the simulated microgravity conditions. MC3T3-E1 were cultured in conventional (CON) and simulated microgravity (SMG), respectively. Then, the expression of the related osteoblastic genes and the specific molecules in NF-κB signaling pathway were measured. The results showed that the mRNA and protein levels of alkaline phosphatase (ALP), osteocalcin (OCN) and type Ⅰ collagen (CoL-Ⅰ) were dramatically decreased under the simulated microgravity. Meanwhile, the NF-κB inhibitor α (IκB-α) protein level was decreased and the expressions of phosphorylation of IκB-α (p-IκB-α), p65 and phosphorylation of p65 (p-p65) were significantly up-regulated in SMG group. In addition, the IL-6 content in SMG group was increased compared to CON. These results indicated that simulated microgravity could activate the NF-κB pathway to regulate MC3T3-E1 cells differentiation.


Assuntos
Diferenciação Celular , NF-kappa B/fisiologia , Transdução de Sinais , Simulação de Ausência de Peso , Células 3T3 , Animais , Camundongos , Osteoblastos
7.
Cancer Sci ; 110(8): 2643-2651, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31222839

RESUMO

Scirrhous-type gastric cancer (SGC) is one of the most intractable cancer subtypes in humans, and its therapeutic targets have been rarely identified to date. Exploration of somatic mutations in the SGC genome with the next-generation sequencers has been hampered by markedly increased fibrous tissues. Thus, SGC cell lines may be useful resources for searching for novel oncogenes. Here we have conducted whole exome sequencing and RNA sequencing on 2 SGC cell lines, OCUM-8 and OCUM-9. Interestingly, most of the mutations thus identified have not been reported. In OCUM-8 cells, a novel CD44-IGF1R fusion gene is discovered, the protein product of which ligates the amino-terminus of CD44 to the transmembrane and tyrosine-kinase domains of IGF1R. Furthermore, both CD44 and IGF1R are markedly amplified in the OCUM-8 genome and abundantly expressed. CD44-IGF1R has a transforming ability, and the suppression of its kinase activity leads to rapid cell death of OCUM-8. To the best of our knowledge, this is the first report describing the transforming activity of IGF1R fusion genes. However, OCUM-9 seems to possess multiple oncogenic events in its genome. In particular, a novel BORCS5-ETV6 fusion gene is identified in the OCUM-9 genome. BORCS5-ETV6 possesses oncogenic activity, and suppression of its message partially inhibits cell growth. Prevalence of these novel fusion genes among SGC awaits further investigation, but we validate the significance of cell lines as appropriate reagents for detailed genomic analyses of SGC.


Assuntos
Adenocarcinoma Esquirroso/genética , Oncogenes/genética , Neoplasias Gástricas/genética , Células 3T3 , Adenocarcinoma Esquirroso/patologia , Animais , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/genética , Células HEK293 , Humanos , Receptores de Hialuronatos/genética , Proteínas de Membrana/genética , Camundongos , Mutação/genética , Receptor IGF Tipo 1/genética , Estômago/patologia , Neoplasias Gástricas/patologia
8.
Biotechnol Appl Biochem ; 66(4): 690-697, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31173404

RESUMO

Osteoporosis is one of the clinical complications of long-term treatment with glucocorticoids (GCs), characterized by systemic damage of bone mass and osteoblast dysfunction. Hydrogen sulfide was found to be involved in GCs-induced osteoblast dysfunction. Osteoblastic MC3T3-E1 cell and mitochondrial function were determined by cell viability, M-CSF level, and ALP activity and superoxide production, membrane potential, and ATP level, respectively. The purpose of this research was to explore the impact of NaHS on osteoblastic MC3T3-E1 cell function as well as on Sirt1 and PGC1α expression in dexamethasone (DEX)-treated osteoblast cells. DEX-treated MC3T3-E1 cells exhibited decreased cell viability and ALP activity, as well as increased M-CSF level; all these changes were dramatically attenuated by NaHS. DEX-treated cells also displayed mitochondrial dysfunction, namely decreased mitochondrial membrane potential and ATP generation and increased superoxide generation, which were partly reversed by NaHS. We confirmed decreased Sirt1 and PGC1α protein expression in DEX-treated MC3T3-E1 cells by Western blot, which was also partly reversed by NaHS. Silencing of Sirt1 abrogated the protective effect of NaHS against DEX-induced cell damage and mitochondrial dysfunction. NaHS alleviates DEX-induced osteoblastic MC3T3-E1 cell injury by improving mitochondrial function.


Assuntos
Dexametasona/farmacologia , Mitocôndrias/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Osteoblastos/patologia , Sulfetos/farmacologia , Células 3T3 , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Camundongos , Mitocôndrias/metabolismo , Osteoblastos/metabolismo
9.
Biochemistry (Mosc) ; 84(6): 686-692, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31238868

RESUMO

Resveratrol has been shown to stimulate differentiation of osteoblastic MC3T3-E1 cells in vitro; however, the mechanisms underlying the anabolic effect of resveratrol on osteoblasts remain largely unknown. Our study was aimed to investigate the molecular mechanism of resveratrol-induced differentiation of MC3T3-E1 cells. MC3T3-E1 cells were treated for 8 days with different concentrations of resveratrol (10-8-10-6 M) and 10-6 M cyclosporine A (CsA), a specific inhibitor of the calcineurin/NFAT pathway. According to the results of pilot studies of cell proliferation and alkaline phosphatase activity, 10-7 M concentration of resveratrol was used in subsequent experiments. The levels of mRNA expression of the osteosis-related genes CaN, NFATc1, and Runx2 were analyzed by real-time RT-PCR; the levels of the corresponding proteins were estimated by Western blot analysis. Resveratrol upregulated expression of the CaN, NFATc1, and Runx2 genes at both mRNA and protein levels compared to the control group (p < 0.05), while CsA reduced the effects of resveratrol (p < 0.05). Using immunohistochemical staining, we showed that resveratrol induced NFATc1 accumulation in the cell nuclei, and treatment with CsA inhibited resveratrol-mediated induction of NFATc1, suggesting that the calcineurin/NFATc1 signaling pathway plays an important role in the regulatory effect of resveratrol on osteoblasts.


Assuntos
Antioxidantes/farmacologia , Calcineurina/metabolismo , Diferenciação Celular/efeitos dos fármacos , Fatores de Transcrição NFATC/metabolismo , Osteoblastos/efeitos dos fármacos , Resveratrol/farmacologia , Células 3T3 , Animais , Calcineurina/genética , Proliferação de Células/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Camundongos , Fatores de Transcrição NFATC/genética , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteogênese/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real
10.
J Colloid Interface Sci ; 551: 72-80, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31075635

RESUMO

Ionic liquid (IL) surfactants have attracted great interest as promising substitutes for conventional surfactants owing to their exceptional and favorable physico-chemical properties. However, most IL surfactants are not eco-friendly and form unstable micelles, even when using a high concentration of the surfactant. In this study, we prepared a series of halogen-free and biocompatible choline-fatty-acid-based ILs with different chain lengths and degrees of saturation, and we then investigated their micellar properties in aqueous solutions. Characterization of the synthesized surface-active ILs (SAILs) was performed by 1H and 13C nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, and elemental analysis. The surface-active properties of the SAILs were investigated by tensiometry, conductometry, and dynamic light scattering measurements. The critical micelle concentration of the SAILs was found to be 2-4 times lower than those of conventional surfactants. The thermodynamic properties of micellization (ΔG0m, ΔH0m, and ΔS0m) indicate that the micellization process of the SAILs is spontaneous, stable, and entropy-driven at room temperature. The cytotoxicity of the SAILs was evaluated using mammalian cell line NIH 3T3. Importantly, [Cho][Ole] shows lower toxicity than the analogous ILs with conventional surfactants. These results clearly suggest that these environmentally friendly SAILs can be used as a potential alternative to conventional ILs for various purposes, including biological applications.


Assuntos
Materiais Biocompatíveis/química , Colina/química , Ácidos Graxos/química , Líquidos Iônicos/química , Tensoativos/química , Células 3T3 , Animais , Sobrevivência Celular/efeitos dos fármacos , Líquidos Iônicos/toxicidade , Camundongos , Micelas , Propriedades de Superfície , Tensoativos/toxicidade , Temperatura Ambiente , Termodinâmica , Água
11.
Phytochemistry ; 164: 111-121, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31125861

RESUMO

An investigation of the n-BuOH fraction of Schnabelia nepetifolia (Benth.) P.D.Cantino led to the isolation and identification of 12 undescribed phenylethanoid glycosides (nepetifosides A-L) and one undescribed phenylmethanoid glycoside (nepetifoside M), together with 23 known compounds. The structures of these compounds were determined by spectroscopic analyses including two-dimensional nuclear-magnetic-resonance (2D-NMR) spectroscopy and chemical-hydrolysis methods. Nepetifoside F exhibited strong activity that significantly increased osteoblast proliferation at three concentrations of 0.1, 1, and 10 µM. Moreover, nepetifoside C and nepetifoside D exhibited moderate activities in promoting the proliferation of osteoblasts at medium and high concentrations of 1 µM and 10 µM, respectively.


Assuntos
Glicosídeos/farmacologia , Lamiaceae/química , Osteoblastos/efeitos dos fármacos , Álcool Feniletílico/farmacologia , Células 3T3 , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Glicosídeos/química , Glicosídeos/isolamento & purificação , Camundongos , Conformação Molecular , Álcool Feniletílico/química , Álcool Feniletílico/isolamento & purificação , Relação Estrutura-Atividade
12.
Cell Biochem Funct ; 37(5): 385-394, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31140646

RESUMO

To explore the molecular mechanism of insulin on proliferation and differentiation of MC3T3-E1 cell under high glucose conditions. We first investigated the effect of different concentrations of insulin on the osteoblast cell proliferation and cell differentiation at various time points by MTT analysis, cell cycle analysis, and expression detection of differentiation genes. Then, we used 200 ng/mL of insulin to treat the osteoblast cell at different time points for identifying the common differentially expressed mRNAs among various time points by RNA sequencing. Thirdly, we performed the gene ontology (GO) and the Kyoto Encyclopaedia of Genes and Genomes (KEGG) analysis to explore the biological function of these common differentially expressed mRNAs. The results showed that insulin promoted the cell proliferation and differentiation of osteoblast cell. In RNA sequencing, a total of 31 common differentially expressed mRNAs were identified between different time points. Mt1, Tmem135, Avp, and Dlg2 were found to be associated with the new bone formation. In addition, three important signalling pathways, namely, lysosome, glutamatergic synapse, and chemokine signalling pathways, were found in the KEGG enrichment analysis. Our work demonstrated that insulin could promote the osteoblast cell proliferation and cell differentiation, which may play a key role in bone formation. SIGNIFICANCE OF THE STUDY: Our result showed that insulin could promote the proliferation and differentiation of osteoblast at both cellular and molecular levels, which may promote the new bone formation in the osteoblasts.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Glucose/metabolismo , Insulina/farmacologia , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Células 3T3 , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Camundongos , Osteoblastos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
13.
Artif Cells Nanomed Biotechnol ; 47(1): 1823-1832, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31066304

RESUMO

Poly(lactic-co-glycolic acid) (PLGA) based biomaterials have many advantages and potential applications in bone tissue engineering. Whereas, a significant problem which could not be ignored was that the acidic by-products generated during its degradation induced severe inflammatory reactions and negatively regulated bone regeneration. In this research, feasibility of using dexamethasone (Dex) to improve the biocompatibility of PLGA is investigated in detail. Hereby, various contents of PLGA/hydroxyapatite (PH) nanofibers loaded with Dex were synthesized by electrospinning technique. It was shown that 0.5% (wt) Dex in PH scaffolds was the minimum content which was required for anti-inflammatory effect. Admittedly, Dex to some extent had cytotoxic effect on osteoblasts and an inhibitory effect on ALP activity in this study; nevertheless, the relatively low Dex (<2% [wt]) had no inhibitory effects on osteoblasts maturation and mineralization. By this token, Dex is a good candidate for improving biocompatibility of PLGA based biomaterials. Moreover, the cytotoxic effects of Dex should be concerned. This study will provide a rationale for optimizing biocompatibility of PLGA based biomaterials by using Dex.


Assuntos
Dexametasona/química , Portadores de Fármacos/química , Durapatita/química , Nanofibras/química , Osteogênese/efeitos dos fármacos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , Células 3T3 , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Liberação Controlada de Fármacos , Teste de Materiais , Camundongos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Ratos , Solventes/química
14.
Eur J Med Chem ; 175: 162-171, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31082763

RESUMO

We have designed novel tropinone-thiazole derivatives that showed high antiproliferative activity against a variety of cancer cell lines via caspase 3/7 activation mechanism. Among the derivatives, compounds 3b-3h were found to exhibit high activity against human leukemia (MV4-11), human lung carcinoma (A549), human breast carcinoma (MCF-7), and skin melanoma (B16-F10) cancer cell lines, with IC50 values of 5.43-11.06 µM. The lead compound 3g increases caspase 3/7 activity in A549 cells 25 times more than the control, and 2 times more than reference drug camptothecin. We have also found that tropinone-thiazole derivatives exhibit high tyrosinase inhibitory activity. The lead compounds 3g and 3h showed tyrosinase inhibition effect, with IC50 values 3.22 and 3.51 µM, respectively. These inhibitory activities are 22 times higher than the activity of kojic acid (IC50 72.27 µM) and 120 times higher than activity of ascorbic acid (IC50 386.5 µM). For compounds 3g and 3h, the experimentally determined lipophilicity correlates very well with their enzymatic activities. These data suggest that presented compounds could constitute lead anticancer drug candidates.


Assuntos
Caspase 3/metabolismo , Caspase 7/farmacologia , Proliferação de Células/efeitos dos fármacos , Descoberta de Drogas , Ativadores de Enzimas/farmacologia , Inibidores Enzimáticos/farmacologia , Monofenol Mono-Oxigenase/antagonistas & inibidores , Tiazóis/química , Tropanos/química , Células 3T3 , Animais , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Caspase 7/química , Linhagem Celular Tumoral , Cromatografia Líquida/métodos , Ensaios de Seleção de Medicamentos Antitumorais , Ativadores de Enzimas/química , Inibidores Enzimáticos/química , Humanos , Concentração Inibidora 50 , Camundongos , Camundongos Endogâmicos BALB C , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Massas por Ionização por Electrospray/métodos , Relação Estrutura-Atividade
15.
Eur J Med Chem ; 176: 50-60, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31096118

RESUMO

New sulfonamides 5/6 derived from 4-methoxyacetophenone 1 were synthesized by N-sulfonation reaction of ammonia (3) and aminopyrimidinone (4) with its sulfonyl chloride derivative 2. Sulfonamides 5 and 6 were used as precursors of two new series of chalcones 8a-f and 9a-f, which were obtained through Claisen-Schmidt condensation with aromatic aldehydes 7a-f. Compounds 5/6, 8a-d, 8f, 9a-d, and 9f were screened by the US National Cancer Institute (NCI) at 10 µM against sixty different human cancer cell lines (one-dose trial). Chalcones 8b and 9b satisfied the pre-determined threshold inhibition criteria and were selected for screening at five different concentrations (100, 10, 1.0, 0.1, and 0.01 µM). Compound 8b exhibited remarkable GI50 values ranging from 0.57 to 12.4 µM, with cytotoxic effects being observed in almost all cases, especially against the cell lines K-562 of Leukemia and LOX IMVI of Melanoma with GI50 = 0.57 and 1.28 µM, respectively. Moreover, all compounds were screened against Mycobacterium tuberculosis H37Rv, chalcones 8a-c and 9a-c were the most active showing MIC values between 14 and 42 µM, and interestingly they were devoid of antibacterial activity against Mycobacterium smegmatis and Staphylococcus aureus. These antituberculosis hits showed however low selectivity, being equally inhibitory to M. tuberculosis and mammalian T3T cells. The chalcone-sulfonamide hybrids 8a-f and 9a-f resulted to be appealing cytotoxic agents with significant antituberculosis activity.


Assuntos
Antineoplásicos/farmacologia , Antituberculosos/farmacologia , Chalconas/farmacologia , Sulfonamidas/farmacologia , Células 3T3 , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/toxicidade , Antituberculosos/síntese química , Antituberculosos/química , Antituberculosos/toxicidade , Linhagem Celular Tumoral , Chalconas/síntese química , Chalconas/química , Chalconas/toxicidade , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Camundongos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Mycobacterium tuberculosis/efeitos dos fármacos , Sulfonamidas/síntese química , Sulfonamidas/química , Sulfonamidas/toxicidade
16.
Nat Commun ; 10(1): 2141, 2019 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-31105267

RESUMO

Costimulation of T cell responses with monoclonal antibody agonists (mAb-AG) targeting 4-1BB showed robust anti-tumor activity in preclinical models, but their clinical development was hampered by low efficacy (Utomilumab) or severe liver toxicity (Urelumab). Here we show that isotype and intrinsic agonistic strength co-determine the efficacy and toxicity of anti-4-1BB mAb-AG. While intrinsically strong agonistic anti-4-1BB can activate 4-1BB in the absence of FcγRs, weak agonistic antibodies rely on FcγRs to activate 4-1BB. All FcγRs can crosslink anti-41BB antibodies to strengthen co-stimulation, but activating FcγR-induced antibody-dependent cell-mediated cytotoxicity compromises anti-tumor immunity by deleting 4-1BB+ cells. This suggests balancing agonistic activity with the strength of FcγR interaction as a strategy to engineer 4-1BB mAb-AG with optimal therapeutic performance. As a proof of this concept, we have developed LVGN6051, a humanized 4-1BB mAb-AG that shows high anti-tumor efficacy in the absence of liver toxicity in a mouse model of cancer immunotherapy.


Assuntos
Anticorpos Monoclonais Humanizados/farmacologia , Imunoterapia/métodos , Melanoma Experimental/terapia , Receptores de IgG/metabolismo , Neoplasias Cutâneas/terapia , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/antagonistas & inibidores , Células 3T3 , Animais , Anticorpos Monoclonais Humanizados/imunologia , Anticorpos Monoclonais Humanizados/uso terapêutico , Linhagem Celular Tumoral/transplante , Ensaios de Seleção de Medicamentos Antitumorais , Técnicas de Introdução de Genes , Humanos , Fígado/efeitos dos fármacos , Melanoma Experimental/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de IgG/genética , Neoplasias Cutâneas/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/genética , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/imunologia
17.
Chem Commun (Camb) ; 55(45): 6429-6432, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-31094377

RESUMO

Concurrently, manipulation of mitochondrial activity and its monitoring have enormous significance in cancer therapy and diagnosis. In this context, a fluorescent probe MitoDP has been developed for validating H2S mediated protonophore (2,4-dinitrophenol, DNP) induced mitochondrial membrane potential change, ROS formation and ATP depletion in cancer cells. The extent of protonophore activation for mitochondrial dysfunction is monitored through fluorescence signalling at 450 nm. The current study provides a proof for the concept of endogenous H2S-mediated controlled and spatial release of bioactive agents, or toxins specifically in mitochondria of cancer cells.


Assuntos
2,4-Dinitrofenol/farmacologia , Corantes Fluorescentes/farmacologia , Sulfeto de Hidrogênio/farmacologia , Mitocôndrias/efeitos dos fármacos , 2,4-Dinitrofenol/química , Células 3T3 , Animais , Proliferação de Células/efeitos dos fármacos , Corantes Fluorescentes/química , Células HCT116 , Células HeLa , Humanos , Sulfeto de Hidrogênio/química , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Mitocôndrias/metabolismo , Estrutura Molecular , Imagem Óptica , Espécies Reativas de Oxigênio/metabolismo , Espectrometria de Fluorescência
18.
Talanta ; 200: 450-457, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31036208

RESUMO

Non enzymatic detection of NADH and H2O2 is of practical significance for both environmental and biological prospective. However, there is no simple, straight forward electrochemical sensor available for sensing of them in real samples. Addressing this challenge, we report a simple stimuli responsive aminophenol, pre-anodized screen printed carbon electrode (SPCE*/AP) based electrochemical probes for dual detection of NADH and H2O2. Aminophenol prepared and adsorbed on the electrode from aminophenylboronic acid via boronic acid deprotection with H2O2. The SPCE*/AP fabricated with this process was characterized by cyclic voltammetry (CV), scanning electron microscope (SEM), Raman spectroscopy, UV-visible spectroscopy, and X-ray photoelectron spectroscopy (XPS). Amperometric detection results showed that SPCE*/AP electrodes exhibited linearity from 50 µM to 500 µM and from 200 µM to 2 mM with a detection limit (S/N = 3) of 4.2 µM and 28.9 µM for NADH and H2O2, respectively. Excellent reproducibility and selectivity for NADH and H2O2 were observed for this electrochemical platform. In addition, the matrix effect was investigated further using the same technique to analyze NADH and H2O2 in human urine samples, human serum samples, cell culture medium (containing 10% fetal bovine serum, FBS), and environmental water samples (tap water and rain water). Also, the present sensor demonstrated promising outcomes with living cells (normal cells and cancer cells).


Assuntos
Aminofenóis/química , Técnicas Eletroquímicas , Corantes Fluorescentes/química , Peróxido de Hidrogênio/análise , NAD/análise , Células 3T3 , Animais , Carbono/química , Linhagem Celular Tumoral , Eletrodos , Humanos , Camundongos
19.
Anal Bioanal Chem ; 411(20): 5223-5231, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31111180

RESUMO

The visualization of subcellular structures is critical for understanding their intracellular function. We prepared two triarylboron-based multinuclear Zn2+ complexes, TAB-1-3Zn2+ and TAB-2-2Zn2+, which can be used as fluorescent probes for nucleoside polyphosphate (NPP) and RNA because their multi Zn2+ center can selectively combine with the phosphate side chain of NPP or RNA, accompanied by a corresponding fluorescence change. Among them, TAB-2-2Zn2+ is more suitable than TAB-1-3Zn2+ for live cell imaging because of its excellent cell membrane permeability resulting from amphiphilicity. Since the various membrane structures in cells are also composed of phosphoric acid bilayers, TAB-2-2Zn2+ may also be used to image various membrane structures. Various colocalization experiments further confirmed that TAB-2-2Zn2+ can achieve clear simultaneous imaging of the cell membrane, the endoplasmic reticulum, and the nucleolus. Graphical abstract.


Assuntos
Compostos de Boro/química , Membrana Celular/metabolismo , Nucléolo Celular/metabolismo , Complexos de Coordenação/química , Retículo Endoplasmático/metabolismo , Corantes Fluorescentes/química , Compostos de Zinco/química , Células 3T3 , Animais , Células HeLa , Células Hep G2 , Humanos , Camundongos , Fótons , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Frações Subcelulares/metabolismo
20.
Med Sci Monit ; 25: 2792-2801, 2019 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-30990212

RESUMO

BACKGROUND The aim of this study was to investigate the significance of folate receptor-mediated staining solution (FRD) in examination of cervical lesions during gynecological examination. MATERIAL AND METHODS A total of 404 patients participated in this study. FRD staining was applied to screen high grade cervical lesions. ThinPrep cytology test (TCT) and human papillomavirus (HPV) testing were also used for screening high grade cervical lesions. Coincidence rate and KAPPA value of different methods were compared by SPSS software. RESULTS As for CIN2+ and CIN3+, sensitivities for HPV testing were (96.92% and 97.78%) >TCT classification 1 (90.77% and 91.11%) >FRD staining (80.00% and 86.67%) >TCT classification 2 (70.77% and 77.78%), respectively. While specificities for HPV testing were (7.08% and 6.44%)

Assuntos
Neoplasia Intraepitelial Cervical/diagnóstico , Detecção Precoce de Câncer/métodos , Neoplasias do Colo do Útero/patologia , Células 3T3 , Adulto , Idoso , Animais , Neoplasia Intraepitelial Cervical/virologia , Colo do Útero/patologia , Feminino , Receptores de Folato com Âncoras de GPI/química , Receptores de Folato com Âncoras de GPI/metabolismo , Ácido Fólico/química , Humanos , Programas de Rastreamento/métodos , Camundongos , Pessoa de Meia-Idade , Papillomaviridae/patogenicidade , Infecções por Papillomavirus/diagnóstico , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal/métodos
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