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1.
Int J Mol Sci ; 25(2)2024 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-38255833

RESUMO

Furanocoumarins are naturally occurring compounds in the plant world, characterized by low molecular weight, simple chemical structure, and high solubility in most organic solvents. Additionally, they have a broad spectrum of activity, and their properties depend on the location and type of attached substituents. Therefore, the aim of our study was to investigate the anticancer activity of furanocoumarins (imperatorin, isoimperatorin, bergapten, and xanthotoxin) in relation to human glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cell lines. The tested compounds were used for the first time in combination with LY294002 (PI3K inhibitor) and sorafenib (Raf inhibitor). Apoptosis, autophagy, and necrosis were identified microscopically after straining with Hoechst 33342, acridine orange, and propidium iodide, respectively. The levels of caspase 3 and Beclin 1 were estimated by immunoblotting and for the blocking of Raf and PI3K kinases, the transfection with specific siRNA was used. The scratch test was used to assess the migration potential of glioma cells. Our studies showed that the anticancer activity of furanocoumarins strictly depended on the presence, type, and location of substituents. The obtained results suggest that achieving higher pro-apoptotic activity is determined by the presence of an isoprenyl moiety at the C8 position of the coumarin skeleton. In both anaplastic astrocytoma and glioblastoma, imperatorin was the most effective in induction apoptosis. Furthermore, the usage of imperatorin, alone and in combination with sorafenib or LY294002, decreased the migratory potential of MOGGCCM and T98G cells.


Assuntos
Astrocitoma , Cromonas , Furocumarinas , Glioblastoma , Glioma , Morfolinas , Humanos , Sorafenibe/farmacologia , Fosfatidilinositol 3-Quinases , Glioma/tratamento farmacológico , Furocumarinas/farmacologia
2.
Int J Mol Sci ; 24(2)2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36674882

RESUMO

We present a method of microstructure fabrication on the tip of the optical fiber using a UV soft-imprint process of polymerizable ionic liquid-based optical resist. Ionic liquid with two UV-sensitive vinylbenzyl groups in the structure was diluted in non-hazardous propylene glycol (PG) to obtain liquid material for imprinting. No additional organic solvent was required. The impact of propylene glycol amount and exposure dose on optical and mechanical properties was investigated. The final procedure of the UV imprint on the optical fiber tip was developed, including the mold preparation, setup building, UV exposure and post-laser cure. As the IL-containing vinylbenzyl groups can also be polymerized by the radical rearrangement of double bonds through thermal heating, the influence of the addition of 1-2% BHT polymerization inhibitor was verified. As a result, we present the fabricated diffraction gratings and the optical fiber spectrometer component-grism (grating-prism), which allows obtaining a dispersion spectrum at the output of an optical in line with the optical fiber long axis, as the main component in an optical fiber spectrometer. The process is very simple due to the fact that its optimization already starts in the process of molecule design, which is part of the trend of sustainable technologies. The final material can be designed by the tailoring of the anion and/or cation molecule, which in turn can lead to a more efficient fabrication procedure and additional functionalities of the final structure.

3.
BMC Plant Biol ; 22(1): 600, 2022 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-36539686

RESUMO

BACKGROUND: Arabinogalactan proteins (AGPs) are extracellular matrix constituents involved in plant response to fungal infection. The aim of the current study was to investigate the antifungal effect of AGPs ex situ and to determine the structural features of AGPs that may have an influence on this activity. The features of AGPs isolated from fruit were investigated with molecular tools based on specific monoclonal antibodies recognizing carbohydrate AGP epitopes. The Antifungal (well-diffusion) Susceptibility Test and the Agar Invasion Test were used to assess the impact of AGPs on Penicillium notatum culture. RESULTS: The results definitely ruled out the influence of AGPs on fungal growth. The immunochemical analyses revealed that AGPs consist mainly of carbohydrate chains composed of ß-linked glucuronosyl residues recognized by LM2 and GlcA-ß(1 → 3)-GalA-α(1 → 2) Rha recognized by JIM13, which do not have the same functional properties outside the plant cell in in vitro experimental conditions. CONCLUSIONS: The action of a single cell wall component does not elicit any influence ex situ. The extensive accumulation of glycan chains of AGPs in infected tissue as a result of a complex mechanism occurring in the cell wall emphasizes the importance of dependencies between particular components of the extracellular matrix in response to fungal attack.


Assuntos
Antifúngicos , Frutas , Frutas/metabolismo , Antifúngicos/metabolismo , Mucoproteínas/metabolismo , Carboidratos , Proteínas de Plantas/metabolismo
4.
Int J Mol Sci ; 22(10)2021 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-34068110

RESUMO

The aim of the study was to investigate the anticancer potential of LY294002 (PI3K inhibitor) and temozolomide using glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cells. Apoptosis, autophagy, necrosis, and granules in the cytoplasm were identified microscopically (fluorescence and electron microscopes). The mitochondrial membrane potential was studied by flow cytometry. The activity of caspases 3, 8, and 9 and Akt was evaluated fluorometrically, while the expression of Beclin 1, PI3K, Akt, mTOR, caspase 12, and Hsp27 was determined by immunoblotting. SiRNA was used to block Hsp27 and PI3K expression. Cell migration and localization of Hsp27 were tested with the wound healing assay and immunocytochemistry, respectively. LY294002 effectively diminished the migratory potential and increased programmed death of T98G and MOGGCCM. Autophagy was dominant in MOGGCCM, while apoptosis was dominant in T98G. LY294002 with temozolomide did not potentiate cell death but redirected autophagy toward apoptosis, which was correlated with ER stress. A similar effect was observed after blocking PI3K expression with siRNA. Transfection with Hsp27 siRNA significantly increased apoptosis related to ER stress. Our results indicate that inhibition of the PI3K/Akt/mTOR pathway sensitizes glioma cells to apoptosis upon temozolomide treatment, which was correlated with ER stress. Hsp27 increases the resistance of glioma cells to cell death upon temozolomide treatment.


Assuntos
Biomarcadores Tumorais/metabolismo , Cromonas/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica , Glioblastoma/tratamento farmacológico , Morfolinas/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Temozolomida/farmacologia , Antineoplásicos Alquilantes/farmacologia , Apoptose , Biomarcadores Tumorais/genética , Proliferação de Células , Inibidores Enzimáticos/farmacologia , Glioblastoma/genética , Glioblastoma/metabolismo , Glioblastoma/patologia , Humanos , Potencial da Membrana Mitocondrial , Necrose , Fosfatidilinositol 3-Quinases/química , Fosfatidilinositol 3-Quinases/genética , Células Tumorais Cultivadas
5.
Molecules ; 25(21)2020 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-33171577

RESUMO

Coumarins, which occur naturally in the plant kingdom, are diverse class of secondary metabolites. With their antiproliferative, chemopreventive and antiangiogenetic properties, they can be used in the treatment of cancer. Their therapeutic potential depends on the type and location of the attachment of substituents to the ring. Therefore, the aim of our study was to investigate the effect of simple coumarins (osthole, umbelliferone, esculin, and 4-hydroxycoumarin) combined with sorafenib (specific inhibitor of Raf (Rapidly Accelerated Fibrosarcoma) kinase) in programmed death induction in human glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cells lines. Osthole and umbelliferone were isolated from fruits: Mutellina purpurea L. and Heracleum leskowii L., respectively, while esculin and 4-hydroxycoumarin were purchased from Sigma Aldrich (St. Louis, MO, USA). Apoptosis, autophagy and necrosis were identified microscopically after straining with specific fluorochromes. The level of caspase 3, Beclin 1, PI3K (Phosphoinositide 3-kinase), and Raf kinases were estimated by immunoblotting. Transfection with specific siRNA (small interfering RNA) was used to block Bcl-2 (B-cell lymphoma 2), Raf, and PI3K expression. Cell migration was tested with the wound healing assay. The present study has shown that all the coumarins eliminated the MOGGCCM and T98G tumor cells mainly via apoptosis and, to a lesser extent, via autophagy. Osthole, which has an isoprenyl moiety, was shown to be the most effective compound. Sorafenib did not change the proapoptotic activity of this coumarin; however, it reduced the level of autophagy. At the molecular level, the induction of apoptosis was associated with a decrease in the expression of PI3K and Raf kinases, whereas an increase in the level of Beclin 1 was observed in the case of autophagy. Inhibition of the expression of this protein by specific siRNA eliminated autophagy. Moreover, the blocking of the expression of Bcl-2 and PI3K significantly increased the level of apoptosis. Osthole and sorafenib successfully inhibited the migration of the MOGGCCM and T98G cells.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Cumarínicos/farmacologia , Glioblastoma/tratamento farmacológico , Magnoliopsida/química , Extratos Vegetais/farmacologia , Sorafenibe/farmacologia , 4-Hidroxicumarinas/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Proteína Beclina-1/genética , Proteína Beclina-1/metabolismo , Caspase 3/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Esculina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Necrose/tratamento farmacológico , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Interferente Pequeno/metabolismo , Umbeliferonas/farmacologia , Quinases raf/metabolismo
6.
Langmuir ; 35(37): 11968-11978, 2019 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-31442379

RESUMO

Planar photonic components can be fabricated with high resolution by electron beam patterning of polymer thin films on solid substrates such as silicon and glass. However, polymer films are normally formed by spin-coating lithographic resists containing not only polymers but also volatile solvents, which is a serious environmental and health issue. Therefore, we investigate a new type of material for planar structure fabrication (i.e., room-temperature ionic liquids (RTILs) with a polymerizable allyl group) that is electron-beam-curable, solvent-free, and thus potentially interesting for processing materials with weak resistance to solvents. We fabricate planar polymer microstructures by electron beam patterning of RTIL thin films in vacuum, which is possible because of the negligible volatility of ionic liquids. Three different polymerizable ionic liquids {i.e., [Allmim][Cl] (1-allyl-3-methylimidazolium chloride), [Allmim][NTf2] (1-allyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide), and [Allmmim][NTf2] (1-allyl-2,3-dimethylimidazolium bis(trifluoromethylsulfonyl)imide)} are compared in terms of the quality of the fabricated microstructures. We demonstrate that the shape of the more viscous RTIL with the Cl- anion is less distorted during electron-beam-activated polymerization than the shape of the less viscous RTILs with a large NTf2- anion. Furthermore, the surface tension of the NTf2-based ionic liquid decreases significantly with temperature as compared to that of the Cl-based ionic liquid. Thus, we suggest that the thermocapillary effect, that is, the Marangoni flow caused by a temperature gradient, might be responsible for the differences in the shape of the RTIL-derived microstructures. Also, we analyze the chemistry of the electron-beam-activated polymerization of RTIL by the use of Fourier-transform infrared spectroscopy (FTIR) and conclude that because of the disappearance of C═C bonds the free radical polymerization is a probable reaction mechanism. Finally, we show that polymerized microstructures are potentially attractive as planar photonic components because of good optical properties such as a high refractive index.

7.
Nanotechnology ; 29(47): 475202, 2018 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-30198858

RESUMO

Electron beam patterning is an important technology in the fabrication of miniaturized photonic devices. The fabrication process conventionally involves the use of radiation sensitive polymer-based solutions (called resists). We propose to replace typical polymer resists with eco-friendly solvent-free room temperature ionic liquids (RTILs), which are polymerized in situ and solidified by an electron beam. It is demonstrated that the shapes of polymerized structures are different for high-viscous Cl-based RTILs and low-viscous NTf2-based RTILs. Due to the the satisfactory quality of the polymerized spatial microstructures and their light transmission properties, the RTIL-derived microstructures are potentially attractive as photonic elements for near-infrared.

8.
Postepy Biochem ; 63(4): 277-286, 2017.
Artigo em Polonês | MEDLINE | ID: mdl-29374429

RESUMO

Neoplasm diseases are one of the main causes of death in Poland and worldwide. Forming and progression of tumour are regulated by the number of factors, among which one of the most important are matrix metalloproteinases (MMPs), zinc-dependant proteases, responsible for remodeling of extracellular matrix (ECM). They may induce cancer progression directly by modifying the ECM, enabling cancer growth and migrating of cells released from tumour, as well as invading adjacent tissue and blood or lymphatic vessels. MMPs may also induce carcinogenesis in indirect way by modifying tumour microenvironment and secreting factors promoting or inhibiting particular processes. There is number of factors secreted by cancer cells, stromal components and ECM elements regulating activation and functionality of matrix metalloproteinases. Understanding the mechanisms and pathways underlying regulation and activation of MMPs is crucial for comprehension of carcinogenesis and metastasis, and may contribute to developing of new therapeutic strategies.


Assuntos
Carcinogênese , Progressão da Doença , Metaloproteinases da Matriz/metabolismo , Metástase Neoplásica , Neoplasias/enzimologia , Neoplasias/patologia , Ativação Enzimática , Matriz Extracelular/metabolismo , Humanos , Neoplasias/tratamento farmacológico
9.
Front Plant Sci ; 15: 1365490, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38571716

RESUMO

Arabinogalactan proteins (AGPs) are proteoglycans with an unusual molecular structure characterised by the presence of a protein part and carbohydrate chains. Their specific properties at different stages of the fruit ripening programme make AGPs unique markers of this process. An important function of AGPs is to co-form an amorphous extracellular matrix in the cell wall-plasma membrane continuum; thus, changes in the structure of these molecules can determine the presence and distribution of other components. The aim of the current work was to characterise the molecular structure and localisation of AGPs during the fruit ripening process in transgenic lines with silencing and overexpression of SlP4H3 genes (prolyl 4 hydroxylase 3). The objective was accomplished through comprehensive and comparative in situ and ex situ analyses of AGPs from the fruit of transgenic lines and wild-type plants at specific stages of ripening. The experiment showed that changes in prolyl 4 hydroxylases (P4H3) activity affected the content of AGPs and the progress in their modifications in the ongoing ripening process. The analysis of the transgenic lines confirmed the presence of AGPs with high molecular weights (120-60 kDa) at all the examined stages, but a changed pattern of the molecular features of AGPs was found in the last ripening stages, compared to WT. In addition to the AGP molecular changes, morphological modifications of fruit tissue and alterations in the spatio-temporal pattern of AGP distribution at the subcellular level were detected in the transgenic lines with the progression of the ripening process. The work highlights the impact of AGPs and their alterations on the fruit cell wall and changes in AGPs associated with the progression of the ripening process.

10.
Cells ; 12(23)2023 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-38067099

RESUMO

BACKGROUND: Gliomas are the most malignant tumors of the central nervous system. One of the factors in their high drug resistance is avoiding programmed death (PCD) induction. This is related to the overexpression of intracellular survival pathways: PI3K-Akt/PKB-mTOR and Ras-Raf-MEK-ERK. Apoptosis and autophagy are co-existing processes due to the interactions between Bcl-2 and beclin-1 proteins. Their complex may be a molecular "toggle-switch" between PCD types. The aim of this research was to investigate the role of Bcl-2:beclin-1 complex in glioma cell elimination through the combined action of LY294002 and sorafenib. METHODS: Drug cytotoxicity was estimated with an MTT test. The type of cell death was evaluated using variant microscopy techniques (fluorochrome staining, immunocytochemistry, and transmission electron microscopy), as well as the Bcl-2:beclin-1 complex formation and protein localization. Molecular analysis of PCD indicators was conducted through immunoblotting, immunoprecipitation, and ELISA testing. SiRNA was used to block Bcl-2 and beclin-1 expression. RESULTS: The results showed the inhibitors used in simultaneous application resulted in Bcl-2:beclin-1 complex formation and apoptosis becoming dominant. This was accompanied by changes in the location of the tested proteins. CONCLUSIONS: "Switching" between apoptosis and autophagy using PI3K and Raf inhibitors with Bcl-2:beclin-1 complex formation opens new therapeutic perspectives against gliomas.


Assuntos
Glioma , Fosfatidilinositol 3-Quinases , Sorafenibe , Humanos , Apoptose , Autofagia , Proteína Beclina-1 , Glioma/tratamento farmacológico , Glioma/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Sorafenibe/farmacologia , Sorafenibe/uso terapêutico
11.
Pest Manag Sci ; 78(2): 446-455, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34505331

RESUMO

BACKGROUND: Development of new plant protection strategies has become an urgent matter in modern agriculture, in view of the evidently proved negative effect of currently used active ingredients of pesticides. In recent years, much effort has been made to eliminate the use of pesticides established to be toxic to pollinators. RESULTS: In this study, we present a group of new bifunctional ionic liquids based on dodine (N-dodecylguanidine) cation whose physical and biological properties have been modified relative to those of the commercially available N-dodecylguanidine acetate. The decreased level of residue of active substances in plant tissues reduces their availability to pollinators, which increases the safety of their use. Moreover, lower environmental impact in combination with high antifungal activity and an additional biological function, that is the systemic acquired resistance induction, are in line with the goals of sustainable agriculture. CONCLUSION: The presented approach shows the possibility of derivatization of commonly used fungicide into the form of bifunctional salts whose physical and biological properties can be easily modified. The paper reports successful design and synthesis of new sustainable and green chemicals for the modern agriculture, being less toxic to the environment and human health but still effective against pathogens. © 2021 Society of Chemical Industry.


Assuntos
Líquidos Iônicos , Praguicidas , Agricultura , Guanidinas , Humanos
12.
Plants (Basel) ; 12(1)2022 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-36616270

RESUMO

Arnica montana L. is one of Europe's endemic endangered medicinal plants, with diverse biological activities commonly used in medicine, pharmacy, and cosmetics. Its flower heads are a rich source of raw material, with antibacterial, antifungal, antiseptic, anti-inflammatory, antiradical, antioxidant, and antitumor properties. The objective of the present study was (i) to characterize the chemical composition of flower heads of A. montana plants cultivated under nitrogen fertilization, (ii) to identify the impact of the nitrogen fertilization and extraction method (water, ethanol) on the antioxidant activity of extracts, and (iii) to determine the role of different nitrogen doses applied during plant cultivation and different extraction methods in the anticancer activity of the extracts through analysis of apoptosis and autophagy induction in HT29, HeLa, and SW620 cell lines. The present study shows that nitrogen is a crucial determinant of the chemical composition of arnica flower heads and the antioxidant and anticancer activity of the analyzed extracts. Nitrogen fertilization can modify the composition of pharmacologically active substances (sesquiterpene lactones, flavonoids, essential oil) in Arnicae flos. The content of sesquiterpene lactones, flavonoids, and essential oil increased with the increase in the nitrogen doses to 60 kg N ha-1 by 0.66%, 1.45%, and 0.27%, respectively. A further increase in the nitrogen dose resulted in a decrease in the content of the analyzed secondary metabolites. Varied levels of nitrogen application can be regarded as a relevant way to modify the chemical composition of arnica flower heads and to increase the anticancer activity, which was confirmed by the increase in the level of apoptosis with the increase in fertilization to a level of 60 kg N ha-1. The fertilization of arnica plants with low doses of nitrogen (30 and 60 kg N ha-1) significantly increased the LOX inhibition ability of the ethanol extracts. The present study is the first report on the anticancer activity of A. montana water extracts, with emphasis on the role of water as a solvent. In further studies of factors modifying the quality of Arnicae flos, attention should be paid to the simultaneous use of nitrogen and other microelements to achieve synergistic results and to the possibility of a more frequent use of water as a solvent in studies on the biological activity of A. montana extracts.

13.
Biomolecules ; 11(8)2021 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-34439756

RESUMO

The present study aimed to demonstrate Lentinus (formerly Pleurotus) sajor-caju (PSC) as a good source of pro-health substances. It has also shown that supplementation of its culture medium with cow milk may further improve its beneficial properties. Intracellular fractions from fungi grown on a medium supplemented with cow milk were analyzed using various biochemical methods for determination of the nutrient composition. Furthermore, anti-cancer properties of selected extracts were investigated on colorectal cancer cell lines (HT-29, LS 180, and SW948) in vitro. Biochemical analysis showed enrichment in health-enhancing compounds, such as proteins or polysaccharides (about 3.5- and 4.5-fold increase in concentration of proteins and carbohydratesin extracts of mycelia cultured on whole milk (PSC2-I), respectively), with a decrease in the level of free radicals (10-fold decrease in extract grown on milk and medium mixture (1:1) (PSC3-II)), which was related to increased catalase and superoxide dismutase activity (7.5-fold increase in catalase activity and 5-fold in SOD activity in PSC3-II compared to the control). Moreover, the viability of the cancer cells was diminished (to 60.0 ± 6.8% and 40.0 ± 8.6% of the control, on HT-29 and SW948 cells, respectively), along with pro-apoptotic (to 18.8 ± 11.8 and 14.7 ± 8.0% towards LS 180 and SW948 cells, respectively) and NO-secreting effects (about 2-fold increase) of the extracts. This study suggests that PSC has multiple nutritional and anti-cancer properties and can be used as a source of healthy biomolecules in modern medicine or functional foods.


Assuntos
Antineoplásicos/farmacologia , Lentinula/metabolismo , Leite/química , Pleurotus/metabolismo , Animais , Antioxidantes/farmacologia , Apoptose , Catalase/metabolismo , Linhagem Celular Tumoral , Células HT29 , Humanos , Necrose , Óxido Nítrico/química , Polissacarídeos/metabolismo , Superóxido Dismutase/metabolismo
14.
Biomolecules ; 11(9)2021 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-34572476

RESUMO

In this study, the influence of two subfractions (with previously proven anti-cancer properties) isolated from wood rot fungus Cerrena unicolor on the formation of a fibrin clot was investigated in the context of potential use as fibrin glue and sealant enhancers and potential wound healing agents. With the use of ROTEM thromboelastometry, we demonstrated that, in the presence of fibrinogen and thrombin, the S6 fraction accelerated the formation of a fibrin clot, had a positive effect on its elasticity modulus, and enhanced the degree of fibrin cross-linking. The S5 fraction alone showed no influence on the fibrin coagulation process; however, in the presence of fibrin, it exhibited a decrease in anti-proliferative properties against the HT-29 line, while it increased the proliferation of cells in general at a concentration of 100 µg/mL. Both fractions retained their proapoptotic properties to a lesser degree. In combination with the S6 fraction in the ratio of 1:1 and 1:3, the fractions contributed to increased inhibition of the activity of matrix metalloproteinases (MMPs). This may suggest anti-metastatic activity of the combined fractions. In conclusion, the potential of the fractions isolated from the C. unicolor secretome to be used as a means of improving the wound healing process was presented. The potential for delivering agents with cytostatic properties introduced far from the site of action or exerting a pro-proliferative effect at the wound site with the aid of a fibrin sealant was demonstrated.


Assuntos
Antineoplásicos/farmacologia , Portadores de Fármacos/química , Adesivo Tecidual de Fibrina/farmacologia , Polyporales/química , Tromboelastografia , Apoptose/efeitos dos fármacos , Coagulação Sanguínea/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Elasticidade , Fibrina/metabolismo , Fungos/efeitos dos fármacos , Gelatina/metabolismo , Humanos , Cinética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Trombina/farmacologia , Viscosidade
15.
Cancers (Basel) ; 13(11)2021 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-34072003

RESUMO

AIM: The anti-glioma effect of lensoside Aß alone and in combination with sorafenib (pro-survival Raf kinase inhibitor) was evaluated for the first time in terms of programmed cell death induction in anaplastic astrocytoma and glioblastoma multiforme cell lines as an experimental model. Apoptosis, autophagy, and necrosis were identified microscopically (fluorescence and scanning microscopes) and confirmed by flow cytometry (mitochondrial membrane potential MMP and cell death). The expression of apoptotic (caspase 3) and autophagic markers (beclin 1) as well as Raf kinase were estimated by immunoblotting. The FTIR method was used to determine the interaction of the studied drugs with lipid and protein groups within cells, while the modes of drug action within the cells were assessed with the FLIM technique. RESULTS: Lensoside Aß itself does not exhibit anti-glioma activity but significantly enhances the anti-cancer potential of sorafenib, initiating mainly apoptosis of up to 90% of cells. It was correlated with an increased level of active caspase 3, a reduced MMP value, and a lower level of Raf kinase. The interaction with membrane structures led to morphological changes typical of programmed death. CONCLUSIONS: Our results indicate that lensoside Aß plays an important role as an adjuvant in chemotherapy with sorafenib and may be a potential candidate in anti-glioma combination therapy.

16.
Fitoterapia ; 142: 104492, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32032635

RESUMO

Furanocoumarins are a class of natural compounds produced by several plants, including those consumed by humans. They have been used medicinally in eastern countries for ages. Given the growing body of evidence about their anticancer potential and observations that naturally occurring compounds potentiate the antitumor activity of chemotherapeutics, more attention is paid to elucidation of the nature of furanocoumarins and the possibility of using thereof in practice. The general mechanism by which furanocoumarins eliminate cancer cells is based on cell cycle blockage and initiation of programmed death like apoptosis or autophagy. The precise molecular mechanism of such an action depends on the chemical structure of furanocoumarins, which is based on the furan ring attached to the coumarin backbone in a linear or angular form as well as the type, location, and number of the substituents attached. The review summarizes the current evidence of the antitumor properties of linear and angular furanocoumarins with special emphasis on the molecular mechanism of elimination of cancer cells via apoptosis and autophagy. Negative aspects of the use of coumarins in anticancer therapy will be also discussed especially in the context of their phototoxicity and potential cancerogenic effect.


Assuntos
Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Furocumarinas/química , Furocumarinas/farmacologia , Humanos , Estrutura Molecular
17.
Sci Rep ; 10(1): 20621, 2020 11 26.
Artigo em Inglês | MEDLINE | ID: mdl-33244134

RESUMO

Arabinogalactan proteins (AGPs) are ubiquitous cell wall and plasma membrane components and are characterised by extensive glycosylation and heterogeneity of their carbohydrate and protein units. The aim of the study was to evaluate the structural features of AGPs present in apple fruits at different stages of the ripening process. AGPs were extracted using the Yariv reagent and examined using SDS-PAGE, immunoblotting, FT-IR, and AFM. In situ analysis, immunofluorescence (CLSM) and immunogold-labelling (TEM), were performed. We demonstrated that AGPs were indeed present in apple fruits at the different stages of the ripening process. The changes in the amount (1.52-2.08 mg g-1), diameter (152.73-75.05 nm), molecular mass (50-250 kDa), and distribution in the cell of AGPs demonstrate their variable presence and changeable structure during the ripening process. We propose specific wavenumbers, i.e. 1265 cm-1, 1117 cm-1, and 960 cm-1, which could be assigned to AGPs. The immunofluorescence and immunogold-labelling results indicate that the JIM13 antibody is the most characteristic for AGPs in apple fruits. This study quantitatively demonstrated for the first time that AGP accumulation occurs in ripe fruits, which is supported by the highest AGPs content, the highest molecular mass, and the appearance of a specific distribution pattern at the cellular level.


Assuntos
Frutas/metabolismo , Mucoproteínas/metabolismo , Parede Celular/metabolismo , Glucosídeos/metabolismo , Malus/metabolismo , Floroglucinol/análogos & derivados , Floroglucinol/metabolismo , Proteínas de Plantas/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier/métodos
18.
Biomed Pharmacother ; 126: 110035, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32113054

RESUMO

Earthworm coelomic fluid (CF) is known as a rich source of various bioactive compounds with promising anticancer features. However, it has been demonstrated that CF affects functionality of both, cancer and normal cells. This non-selective activity causes a major problem for medical application of CF. In this study, we present the anticancer activity of the active protein-carbohydrate fraction (AF) isolated from thermally treated CF of earthworm Dendrobaena veneta. The in vitro effect of the AF was examined in human colon model including normal human colon epithelium (CCD 841 CoTr) and human colon adenocarcinoma (HT-29 and LS180) cell lines. We investigated the impact of AF on cell viability neutral red and lactate dehydrogenase assays, morphology May-Grünwald-Giemsa staining assay proliferation MTT tetrazolium salt and BrdU incorporation assays as well as cell cycle progression propidium iodide/RNase staining and the activity of human 20S proteasome the hydrolysis of AMC from a Suc-LLVY-AMC peptide substrate. Additionally, the influence of AF on apoptosis was examined in HT-29 cells by Annexin V/PI, Hoechst 33342 staining and active caspase-3 assays. Our investigation demonstrated that AF at the tested concentration range does not affect the viability and morphology of CCD 841 CoTr cells. Simultaneously, AF inhibits human 20S proteasome activity as well as significantly decreases mitochondrial metabolism, disturbs cell cycle and induces apoptosis via activation of procaspase-3 in HT-29 cancer cells. Obtained results demonstrate the antiproliferative and proapoptotic activity of AF that can be useful in developing therapeutic strategies to treat human colon cancer.


Assuntos
Adenocarcinoma/tratamento farmacológico , Apoptose/efeitos dos fármacos , Líquidos Corporais/química , Carboidratos/farmacologia , Oligoquetos/metabolismo , Proteínas/farmacologia , Animais , Carboidratos/química , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Células HT29 , Humanos , Proteínas/química
19.
Eur J Pharmacol ; 881: 173207, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32446712

RESUMO

In the recent years, coumarin bioactive compounds have been identified to posess anticancer properties. Therefore, the aim of the present study was to investigate for the first time the efficacy of osthole, umbelliferone, esculin, and 4-hydroxycoumarin, alone and in combination with Temozolomide, in the elimination of deadly brain tumors, anaplastic astrocytoma (AA) and glioblastoma multiforme (GBM) cells via programmed death. Our results indicated that osthole, umbelliferone, esculin, and 4-hydroxycoumarin initiated mainly apoptosis in the T98G and MOGGCCM cells. Osthole was the most effective. It also initiated autophagy in a small percentage of the cell population. The co-incubation with Temozolomide did not increase the pro-apoptotic potential of natural compounds but decreased the level of autophagy in the T98G cells. Apoptosis was associated with reduced mitochondrial membrane potential, activation of caspase 3, inhibition of Bcl-2 expression and the presence of a Bcl-2/Beclin 1. Blocking of Bcl-2 expression resulted in promotion of apoptosis, but not autophagy, in the MOGGCCM and T98G lines. It also sensitized astrocytoma cells, but not GBM, to the combined osthole and TMZ treatment, which was correlated with a reduced level of Beclin 1 and increased expression of caspase 3. Osthole and TMZ, alone and in combination, inhibited the migratory phenotype of the GBM and AA cells. In summary, our results indicated that osthole effectively eliminated glioma cells via apoptosis, what was correlated with Bcl-2/Beclin 1 complex formation. Considering the anti-migratory effect, osthole and Temozolomide display antiglioma potential but it needs further extensive studies.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Cumarínicos/farmacologia , Glioma/tratamento farmacológico , Temozolomida/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Proteína Beclina-1/genética , Proteína Beclina-1/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Necrose , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais
20.
ChemistryOpen ; 8(7): 972-983, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31360624

RESUMO

A new synthetic approach and full spectral (NMR, IR, MS) and ion chromatographic characterization (IC) of nitrogen-based ionic liquids bearing allyl- or ethyl- substituent and triflate, tosylate, methyl sulfate or methanesulfonate anion has been presented. On a sample of 16 new ionic liquids, the versatility of the anion exchange method has been proven. In the metathesis reactions that have been carried out, the halide anion was exchanged in ionic liquid with an alkyl sulfonate based anion using alkylating agents. The results obtained using ion chromatographic analysis on the newly synthesized compounds have been discussed. Also, the utilization of a gaseous methyl halide by-product, obtained in the metathesis reaction and otherwise difficult to synthesize, has been presented. This approach ensured high atom economy of the overall process, which makes the proposed methodology sustainable and eco-friendly.

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