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1.
Microb Pathog ; 189: 106576, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38382626

RESUMO

Serratia marcescens is commonly noted to be an opportunistic pathogen and is often associated with nosocomial infections. In addition to its high antibiotic resistance, it exhibits a wide range of virulence factors that confer pathogenicity. Targeting quorum sensing (QS) presents a potential therapeutic strategy for treating bacterial infections caused by S. marcescens, as it regulates the expression of various virulence factors. Inhibiting QS can effectively neutralize S. marcescens' bacterial virulence without exerting stress on bacterial growth, facilitating bacterial eradication by the immune system. In this study, the antibacterial and anti-virulence properties of eugenol against Serratia sp. were investigated. Eugenol exhibited inhibitory effects on the growth of Serratia, with a minimal inhibitory concentration (MIC) value of 16.15 mM. At sub-inhibitory concentrations, eugenol also demonstrated antiadhesive and eradication activities by inhibiting biofilm formation. Furthermore, it reduced prodigiosin production and completely inhibited protease production. Additionally, eugenol effectively decreased swimming and swarming motilities in Serratia sp. This study demonstrated through molecular modeling, docking and molecular dynamic that eugenol inhibited biofilm formation and virulence factor production in Serratia by binding to the SmaR receptor and blocking the formation of the HSL-SmaR complex. The binding of eugenol to SmaR modulates biofilm formation and virulence factor production by Serratia sp. These findings highlight the potential of eugenol as a promising agent to combat S. marcescens infections by targeting its virulence factors through quorum sensing inhibition.


Assuntos
Percepção de Quorum , Serratia , Biofilmes , Eugenol/farmacologia , Serratia marcescens , Fatores de Virulência/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismo
2.
Molecules ; 28(9)2023 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-37175327

RESUMO

A series of new [1,2,4]triazolo[4,3-a]pyrimidine derivatives was prepared using a one-pot three-component synthesis from 5-amino-1-phenyl-1H-1,2,4-triazoles, aromatic aldehydes and ethyl acetoacetate. The compound structures were confirmed by IR, 1H-NMR, 13C-NMR, HRMS and X-ray analyses. The biological activity of these compounds as antitumor agents was evaluated. Their antitumor activities against cancer cell lines (MDA-MB-231 and MCF-7) were tested by the MTT in vitro method. Among them, compounds 4c and 4j displayed the best antitumor activity with IC50 values of 17.83 µM and 19.73 µM against MDA-MB-231 and MCF-7 cell lines, respectively, compared to the Cisplatin reference.


Assuntos
Antineoplásicos , Pirimidinas , Humanos , Pirimidinas/química , Antineoplásicos/química , Cisplatino/farmacologia , Células MCF-7 , Espectroscopia de Ressonância Magnética , Ensaios de Seleção de Medicamentos Antitumorais , Relação Estrutura-Atividade , Proliferação de Células , Linhagem Celular Tumoral , Estrutura Molecular
3.
Microb Pathog ; 164: 105449, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35183700

RESUMO

The aim of this work was to produce a red tripyrrole pigment prodigiosin from Serratia sp. C6LB strain, to investigate the promising antimicrobial properties on Gram-positive and Gram-negative bacterial strains. The research was also proposed to evaluate the antibiofilm activity on Staphylococcus epidermidis S61 biofilm and its cytotoxic activity against human cancer cell lines. The production and structural elucidation of prodigiosin was carried out using spectrophotometric scanning, TLC, HPLC, FTIR and NMR analysis. The pigment production was optimized using mannose and peptone as carbon and nitrogen sources, respectively. The study confirmed promising antibacterial properties of prodigiosin on eight Gram-positive and Gram-negative bacterial strains with MICs values ranged from 0.039 to 2.5 mg/mL. Antiadhesive activity test of prodigiosin on Staphylococcus epidermidis S61 biofilm exhibited 99.9% inhibition, whereas maximum biofilm eradication activity reached 65%. Cytotoxic activity showed IC50 of 16 µg/mL and 6.7 µg/mL against breast cancer lines MCF-7 and MDA-MB231, respectively.


Assuntos
Prodigiosina , Serratia , Animais , Antibacterianos/metabolismo , Biofilmes , Humanos , Leite , Serratia marcescens
4.
Molecules ; 27(22)2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36431976

RESUMO

Ten new differently substituted 3-benzyl-5-aryl-3,5-dihydro-4H-benzo[6,7]chromeno[2,3-d]pyrimidin-4,6,11-triones 3 were synthesized by a simple and cost-efficient procedure in a one-pot, three-component reaction from readily available ethyl 2-amino-4-aryl-5,10-dioxo-5,10-dihydro-4H-benzo[g]chromene-3-carboxylates, benzylamine and triethyl orthoformate under solvent- and catalyst-free conditions. All the new compounds were screened for their antiproliferative activity against two colorectal-cancer-cell lines. The results showed that the compounds 3-benzyl-5-phenyl-3,5-dihydro-4H-benzo[6,7]chromeno[2,3-d]pyrimidine-4,6,11-trione (3a) and 3-benzyl-5-(3-hydroxyphenyl)-3,5-dihydro-4H-benzo[6,7]chromeno[2,3-d]pyrimidine-4,6,11-trione (3g) exhibited the most potent balanced inhibitory activity against human LoVo and HCT-116 cancer cells.


Assuntos
Neoplasias Colorretais , Pirimidinas , Humanos , Pirimidinas/química , Células HCT116 , Benzopiranos/química , Neoplasias Colorretais/tratamento farmacológico
5.
J Cell Physiol ; 236(7): 4997-5011, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33305427

RESUMO

The epidermal growth factor receptor (EGFR) harbors a calmodulin (CaM)-binding domain (CaM-BD) and a CaM-like domain (CaM-LD) upstream and downstream, respectively, of the tyrosine kinase (TK) domain. We demonstrate in this paper that deletion of the positively charged CaM-BD (EGFR/CaM-BD∆) inactivated the TK activity of the receptor. Moreover, deletion of the negatively charged CaM-LD (EGFR/CaM-LD∆), leaving a single negative residue (glutamate), reduced the activity of the receptor. In contrast, substituting the CaM-LD with a histidine/valine-rich peptide (EGFR/InvCaM-LD) caused full inactivation. We also demonstrated using confocal microscopy and flow cytometry that the chimera EGFR-green fluorescent protein (GFP)/CaM-BD∆, the EGFR/CaM-LD∆, and EGFR/InvCaM-LD mutants all bind tetramethylrhodamine-labelled EGF. These EGFR mutants were localized at the plasma membrane as the wild-type receptor does. However, only the EGFR/CaM-LD∆ and EGFR/InvCaM-LD mutants appear to undergo ligand-dependent internalization, while the EGFR-GFP/CaM-BD∆ mutant seems to be deficient in this regard. The obtained results and in silico modelling studies of the asymmetric structure of the EGFR kinase dimer support a role of a CaM-BD/CaM-LD electrostatic interaction in the allosteric activation of the EGFR TK.


Assuntos
Calmodulina/metabolismo , Membrana Celular/metabolismo , Animais , Células CHO , Sinalização do Cálcio/fisiologia , Linhagem Celular , Cricetulus , Ativação Enzimática/fisiologia , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/genética , Receptores ErbB/metabolismo , Humanos , Ligação Proteica/fisiologia , Domínios Proteicos/fisiologia , Proteínas Tirosina Quinases/metabolismo
6.
Biochem Genet ; 58(1): 102-128, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31286319

RESUMO

Plant NHX antiporters are critical for cellular pH, Na+, and K+ homeostasis and salt tolerance. Even though their genomic and functional studies have been conducted in many species, the grapevine NHX family has not been described yet. Our work highlights the presence of six VvNHX genes whose phylogenetic analysis revealed their classification in two distinct groups: group I vacuolar (VvNHX1-5) and group II endosomal (VvNHX6). Several cis-acting regulatory elements related to tissue-specific expression, transcription factor binding, abiotic/biotic stresses response, and light regulation elements were identified in their promoter. Expression profile analyses of VvNHX genes showed variable transcription within organs and tissues with diverse patterns according to biochemical, environmental, and biotic treatments. All VvNHXs are involved in berry growth, except VvNHX5 that seems to be rather implicated in seed maturation. VvNHX4 would be more involved in floral development, while VvNHX2 and 3 display redundant roles. QPCR expression analyses of VvNHX1 showed its induction by NaCl and KNO3 treatments, whereas VvNHX6 was induced by ABA application and strongly repressed by PEG treatment. VvNHX1 plays a crucial role in a bunch of grape developmental steps and adaptation responses through mechanisms of phyto-hormonal signaling. Overall, VvNHX family members could be valuable candidate genes for grapevine improvement.


Assuntos
Desenvolvimento Vegetal/genética , Proteínas de Plantas/genética , Trocadores de Sódio-Hidrogênio/genética , Estresse Fisiológico/genética , Vitis/crescimento & desenvolvimento , Vitis/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas
7.
Ecotoxicol Environ Saf ; 190: 110103, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31887707

RESUMO

Multi-contaminated industrial wastewaters pose serious environmental risks due to high toxicity and non-biodegradability. The work reported here evaluated the ability of Pseudomonas aeruginosa strain Gb30 isolated from desert soil to simultaneously remove cadmium (Cd) and Reactive Black 5 (RB5), both common contaminants in various industrial effluents. The strain was able to grow normally and decolorize 50 mg L-1 RB5 within 24 h of incubation in the presence of 0.629 m mol L-1 of Cd2+. In order to evaluate strain performance in RB5 detoxification, a cytotoxicity test using Human Embryonic Kidney cells (HEK293) was used. Cadmium removal from culture media was determined using atomic adsorption. Even in presence of (0.115 + 0.157 + 0.401 + 0.381) m mol L-1, respectively, of Cr6+, Cd2+, Cu2+ and Zn2+ in the growth medium, strain Gb30 successfully removed 35% of RB5 and 44%, 36%, 59% and 97%, respectively, of introduced Zn2+, Cu2+, Cr6+ and Cd2+, simultaneously. In order to understand the mechanism of Cd removal used by P. aeruginosa strain Gb30, biosorption and bioaccumulation abilities were examined. The strain was preferentially biosorbing Cd on the cell surface, as opposed to intracellular bioaccumulation. Microscopic investigations using AFM, SEM and FTIR analysis of the bacterial biomass confirmed the presence of various structural features, which enabled the strain to interact with metal ions. The study suggests that Pseudomonas aeruginosa Gb30 is a potential candidate for bioremediation of textile effluents in the presence of complex dye-metal contamination.


Assuntos
Biodegradação Ambiental , Cádmio/metabolismo , Naftalenossulfonatos/metabolismo , Poluentes do Solo/metabolismo , Adsorção , Bactérias/metabolismo , Biomassa , Células HEK293 , Humanos , Metais Pesados/análise , Pseudomonas aeruginosa/metabolismo , Solo , Poluentes do Solo/análise , Águas Residuárias/química
8.
Biodegradation ; 30(4): 287-300, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30600423

RESUMO

The bacterial strain F4, isolated from olive oil-contaminated soil, has been found to produce biosurfactants as confirmed by oil displacement test and the emulsification index results. The identification of the strain F4, by 16S ribosomal RNA gene, showed a close similarity to Bacillus safensis, therefore the strain has been termed Bacillus safensis F4. The Thin Layer Chromatography (TLC) and the High Pressure Liquid Chromatography-Mass Spectrometry (HPLC-MS/MS) demonstrated that the biosurfactant had a lipopeptide structure and was classified as surfactin. The present study showed also that the produced biosurfactant has an important antibacterial activity against several pathogen strains as monitored with minimum inhibitory concentration (MIC) micro-assays. In particular, it presented an interesting anti-planktonic activity with a MIC of 6.25 mg mL-1 and anti-adhesive activity which exceeded 80% against the biofilm-forming Staphylococcus epidermidis S61 strain. Moreover, the produced lipopeptide showed an antitumor activity against T47D breast cancer cells and B16F10 mouse melanoma cells with IC50 of 0.66 mg mL-1 and 1.17 mg mL-1, respectively. Thus, our results demonstrated that Bacillus safensis F4 biosurfactant exhibited a polyvalent activity via a considerable antibiofilm and antitumoral potencies.


Assuntos
Bacillus , Animais , Antibacterianos , Biodegradação Ambiental , Camundongos , Tensoativos , Espectrometria de Massas em Tandem
9.
Lipids Health Dis ; 16(1): 190, 2017 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-28969677

RESUMO

BACKGROUND: Rosmarinus officinalis L. from Tunisia, popularly known as rosemary, is of a considerable importance for its medicinal uses and aromatic value. The aim of this study was to examine the chemical composition of Rosmarinus officinalis essential oil (ROEO) and to evaluate its antibiofilm activity on biofilm-forming bacterium and its anticancer activity on cancer cell lines. METHODS: The chemical composition of Rosmarinus officinalis essential oil (ROEO) was analyzed by GC-MS and its antibacterial activity was evaluated by micro-dilution method. The antibofilm activity of ROEO was evaluated using the crystal violet test and the cytotoxicity activity was determined by the MTT assay. RESULTS: In this research, thirty-six compounds were identified in ROEO using GC-MS analyses. The main components were 1,8-cineole (23.56%), camphene (12.78%), camphor (12.55%) and ß-pinene (12.3%). The antibacterial activity of ROEO was evaluated by micro-dilution method. The oil exhibited inhibition and bactericidal effect against two strains: Staphylococcus aureus ATCC 9144 and Staphylococcus epidermidis S61. It was found that the minimum inhibitory concentration (MIC) obtained for S. aureus and S. epidermidis ranged from 1.25 to 2.5 and from 0.312 to 0.625 µl ml-1, respectively and the minimum bactericidal concentration (MBC) were in the order of 5 and 2.5 µl ml-1, respectively. Furthermore, this oil showed a S. epidermidis biofilm inhibition more than 57% at a concentration of 25 µl ml-1. The eradication of 67% of the established biofilm was observed at a concentration of 50 µl ml-1 of ROEO, whereas the dose of 25 µl ml-1 removed only 38% of preformed biofilm. ROEO strongly inhibited the proliferation of Hela and MCF-7 cells with IC50 values of 0.011 and 0.253 µl ml-1, respectively. CONCLUSION: Our results demonstrate that ROEO could have a potential role in the treatment of diseases related to infection by microorganisms or proliferation of cancer cells.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Óleos Voláteis/farmacologia , Rosmarinus/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Monoterpenos Bicíclicos , Biofilmes/crescimento & desenvolvimento , Compostos Bicíclicos com Pontes/isolamento & purificação , Cânfora/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Monoterpenos Cicloexânicos , Cicloexanóis/isolamento & purificação , Eucaliptol , Cromatografia Gasosa-Espectrometria de Massas , Células HeLa , Humanos , Células MCF-7 , Testes de Sensibilidade Microbiana , Monoterpenos/isolamento & purificação , Óleos Voláteis/química , Óleos Voláteis/isolamento & purificação , Plantas Medicinais , Sesquiterpenos Policíclicos , Sesquiterpenos/isolamento & purificação , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus epidermidis/crescimento & desenvolvimento , Terpenos/isolamento & purificação , Tunísia
10.
J Recept Signal Transduct Res ; 36(1): 21-5, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-25343691

RESUMO

UNLABELLED: The use of some classic antibiotics was recently shown to inhibit growth and to induce apoptosis in human LOVO colon cancer cells. In this study, we describe that ciprofloxacin (CI), trimebutine maleate (COL) and tiemonium methylsulfate (VIS) greatly inhibit cell proliferation in vitro. Proliferation inhibition reached its maximum at 10(-4 )M, 10(-3 )M and 10(-2 )M, respectively, for COL, CI and VIS. Moreover, phospho-extracellular-regulated kinase was totally abrogated in non-apoptotic cytotoxicity of VIS but decreases or increases in the apoptotic inhibition, respectively, of COL and CI treatments. ABBREVIATIONS: CI: ciprofloxacin; COL: trimebutine maleate; VIS: tiemonium methylsulfate; MAPK/Erk: mitogen-activated protein kinases/extracellular-regulated kinase.


Assuntos
Ciprofloxacina/farmacologia , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Morfolinas/farmacologia , Trimebutina/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Ativação Enzimática , Fármacos Gastrointestinais/farmacologia , Humanos , Inibidores da Topoisomerase II/farmacologia , Células Tumorais Cultivadas
11.
J Recept Signal Transduct Res ; 35(2): 202-6, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25319474

RESUMO

The mitogen-activated protein kinase (MAPK) signaling pathway plays key roles in the transmission of proliferative signals in normal and dysregulated cells. Nevertheless, some studies have shown that activation of the extracellular regulated kinases 1/2 (Erk1/2) is involved in apoptosis. In this study, we evaluate the effect of two fertilizing drugs, clomiphene citrate and estradiol, on the activation of Erk1/2 and the viability of two breast cancer cell lines, MCF-7 (hormone dependent) and BT20 (hormone independent).We show that both drugs induce Erk1/2 phosphorylation in MCF-7 and BT20 cells despite their opposite effect on cell viability. In fact, clomiphene citrate is significantly proapoptotic while estradiol promotes cell proliferation. The fact that phospho-Erk1/2 is a common element to both mechanisms suggests that specific factors deciding between proliferation and apoptosis must be operative downstream of this signaling pathway.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/biossíntese , Proteína Quinase 3 Ativada por Mitógeno/biossíntese , Apoptose/efeitos dos fármacos , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Sobrevivência Celular/efeitos dos fármacos , Clomifeno/administração & dosagem , Estradiol/administração & dosagem , Feminino , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células MCF-7 , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/genética , Fosforilação/efeitos dos fármacos
12.
Chem Biodivers ; 12(5): 733-42, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-26010662

RESUMO

Cumin (Cuminum cyminum L.) is a small annual and herbaceous plant belonging to the Apiaceae family. It is a multipurpose plant species cultivated in the Middle East, India, China, and several Mediterranean countries, including Tunisia. Its fruit, known as cumin seed, is most widely used for culinary and medicinal purposes. It is generally used as a food additive, popular spice, and flavoring agent in many cuisines. Cumin has also been widely used in traditional medicine to treat a variety of diseases, including hypolipidemia, cancer, and diabetes. The literature presents ample evidence for the biological and biomedical activities of cumin, which have generally been ascribed to its content and action of its active constituents, such as terpens, phenols, and flavonoids. The present paper provides an overview of phytochemical profile, biological activities, and ethnomedical and pharmacological uses of Cumin.


Assuntos
Pesquisa Biomédica , Cuminum , Medicina Tradicional , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Cuminum/química , Estrutura Molecular , Extratos Vegetais/química
13.
ScientificWorldJournal ; 2014: 830923, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25389535

RESUMO

Our success in producing an active epidermal growth factor receptor (EGFR) tyrosine kinase in Escherichia coli encouraged us to express the full-length receptor in the same host. Despite its large size, we were successful at producing the full-length EGFR protein fused to glutathione S-transferase (GST) that was detected by Western blot analysis. Moreover, we obtained a majoritarian truncated GST-EGFR form detectable by gel electrophoresis and Western blot. This truncated protein was purified and confirmed by MALDI-TOF/TOF analysis to belong to the N-terminal extracellular region of the EGFR fused to GST. Northern blot analysis showed two transcripts suggesting the occurrence of a transcriptional arrest.


Assuntos
Processamento Alternativo , Códon sem Sentido , Receptores ErbB/genética , Proteínas Recombinantes de Fusão/genética , Sequência de Aminoácidos , Clonagem Molecular , Receptores ErbB/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Humanos , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão/metabolismo , Transcrição Gênica
14.
Enzyme Microb Technol ; 180: 110477, 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-39003969

RESUMO

The present paper deals with the preparation and annotation of a surfactin(s) derived from a culture of the endophytic bacterium Bacillus 15 F. The LC-MS analysis of the acetonitrile fraction confirmed the presence of surfactins Leu/Ile7 C15, Leu/Ile7 C14 and Leu/Ile7 C13 with [M+H]+ at m/z 1036.6895, 1022.6741 and 1008.6581, respectively. Various concentrations of the surfactin(s) (hereafter referred to as surfactin-15 F) were used to reduce the adhesion of Staphylococcus epidermidis S61, which served as a model for studying antibiofilm activity on polystyrene surfaces. Incubation of Staphylococcus epidermidis S61 with 62.5 µg/ml of surfactin-15 F resulted in almost complete inhibition of biofilm formation (90.3 ± 3.33 %), and a significant reduction of cell viability (resazurin-based fluorescence was more than 200 times lower). The antiadhesive effect of surfactin-15 F was confirmed by scanning electron microscopy. Surfactin-15 F demonstrated an eradication effect against preformed biofilm, causing severe disruption of Staphylococcus epidermidis S61 biofilm structure and reducing viability. The results suggest that surfactins produced by endophytic bacteria could be an alternative to synthetic products. Surfactin-15 F, used in wound dressings, demonstrated an efficient treatment of the preformed Staphylococcus epidermidis S61 biofilm, and thus having a great potential in medical applications.

15.
Bone Joint Res ; 13(2): 83-90, 2024 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-38368904

RESUMO

Aims: The present study investigated receptor activator of nuclear factor kappa-Β ligand (RANKL), osteoprotegerin (OPG), and Runt-related transcription factor 2 (RUNX2) gene expressions in giant cell tumour of bone (GCTB) patients in relationship with tumour recurrence. We also aimed to investigate the influence of CpG methylation on the transcriptional levels of RANKL and OPG. Methods: A total of 32 GCTB tissue samples were analyzed, and the expression of RANKL, OPG, and RUNX2 was evaluated by quantitative polymerase chain reaction (qPCR). The methylation status of RANKL and OPG was also evaluated by quantitative methylation-specific polymerase chain reaction (qMSP). Results: We found that RANKL and RUNX2 gene expression was upregulated more in recurrent than in non-recurrent GCTB tissues, while OPG gene expression was downregulated more in recurrent than in non-recurrent GCTB tissues. Additionally, we proved that changes in DNA methylation contribute to upregulating the expression of RANKL and downregulating the expression of OPG, which are critical for bone homeostasis and GCTB development. Conclusion: Our results suggest that the overexpression of RANKL/RUNX2 and the lower expression of OPG are associated with recurrence in GCTB patients.

16.
ScientificWorldJournal ; 2013: 807284, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24187524

RESUMO

The present work concerns the heterologous expression of the intracellular domain harbouring the tyrosine kinase activity of the epidermal growth factor receptor (EGFR). Protein expression was improved thanks to the deletion of a 13-amino acid peptide of the juxtamembrane region (JM). The recombinant proteins were produced as a glutathione S-transferase (GST) fusion in Escherichia coli, and the solubilisation was performed by sarkosyl addition during extraction. The produced proteins spontaneously dimerize allowing the activation of the tyrosine kinase domain in the presence of [γ-(32)P]ATP. The activity assay has revealed the autophosphorylation of EGFR proteins which was decreased in the presence of genistein. Our system could facilitate the screening of EGFR inhibitors without the need of adding an exogenous substrate.


Assuntos
Receptores ErbB/metabolismo , Escherichia coli/metabolismo , Trifosfato de Adenosina/metabolismo , Reagentes de Ligações Cruzadas/farmacologia , Dimerização , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/química , Receptores ErbB/isolamento & purificação , Regulação Enzimológica da Expressão Gênica , Genes erbB-1 , Genisteína/farmacologia , Glutaral/farmacologia , Glutationa Transferase/genética , Humanos , Fosforilação , Fosfotirosina/análise , Inibidores de Proteínas Quinases/farmacologia , Processamento de Proteína Pós-Traducional , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Sarcosina/análogos & derivados , Deleção de Sequência , Solventes
17.
Med Oncol ; 40(11): 309, 2023 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-37773302

RESUMO

In this study, we conducted a comprehensive assessment of the cytotoxicity of three glucocorticoids, namely Hydrocortisone, Dexamethasone, and Methylprednisolone, using three different human cell lines: MDA-MB-231, MCF-7 (both adenocarcinoma cell lines), and HEK293 (kidney epithelial cell line). At lower concentrations exceeding 50 µM, we did not observe any significant toxic effects of these glucocorticoids. However, when exposed to higher concentrations, Hydrocortisone exhibited dose-dependent cytotoxic effects on all three cell lines, with calculated IC50 values of 12 ± 0.6 mM for HEK293, 2.11 ± 0.05 mM for MDA-MB-231, and 2.73 ± 0.128 mM for MCF-7 cells after 48 h of exposure. Notably, Hydrocortisone, at its respective IC50 concentrations, demonstrated an inhibitory effect on the proliferation of the cancer cell lines, as evidenced by a substantial reduction in BrdU absorbance in a dose-dependent manner, coupled with a markedly reduced rate of colony formation in treated cells. Furthermore, Hydrocortisone exhibited remarkable anti-migratory properties in MDA-MB-231 and MCF-7 cells, and it induced cell cycle arrest in the SubG1 phase in MDA-MB-231 cells. In addition to these effects, Hydrocortisone triggered apoptosis in both cancer cell types, leading to observable morphological changes. This apoptotic response was characterized by a significant increase in the activity of caspase-3, which was time-dependent. Additionally, Hydrocortisone downregulated the expression of anti-apoptotic Bcl-2 proteins. In summary, our findings underscore the safety of clinical doses in terms of cell toxicity meanwhile increased concentration were showing an anti-proliferative potential of Hydrocortisone, particularly against adenocarcinoma breast cancer cell lines.


Assuntos
Adenocarcinoma , Antineoplásicos , Neoplasias da Mama , Humanos , Feminino , Células MCF-7 , Linhagem Celular Tumoral , Glucocorticoides/farmacologia , Hidrocortisona/farmacologia , Neoplasias da Mama/tratamento farmacológico , Células HEK293 , Antineoplásicos/farmacologia , Apoptose , Rim , Proliferação de Células
18.
J Recept Signal Transduct Res ; 32(3): 142-9, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22394363

RESUMO

BACKGROUND: The prognosis of breast carcinoma is related to a large variety of clinical and pathological factors. Currently, only oestrogen (ER) and progesterone (PR) receptors and human epidermal growth factor receptor 2 (HER2) are used in routine pathological assessment as biomarkers. The aim of this study was to evaluate the prognostic impact of epidermal growth factor receptor (EGFR) expression individually and in combination to classical biomarkers (HER2, ER, and PR), and its relation to tumors with triple negative profile in Tunisian breast carcinoma. METHODS: Immunohistochemistry was used to estimate the rate expression of these receptors. Univariate and multivariate analyses were used to explore the prognostic significance of EGFR in this study. RESULTS: The expression rate of EGFR was 28.6%. EGFR expression was inversely correlated to that of ER (P < 0.001). Significant correlations between the expression of EGFR and the high histological Scarff-Bloom-Richardson (SBR) grade (P = 0.038) and also with tumors size (P = 0.041) were observed. The triple negative profile (TN: ER-/PR-/HER2-) was present in 17.3% of cases. EGFR overexpression was positively associated with this clinical aggressive profile (P < 0.001). Survival analysis showed that EGFR expression was associated with poor survival of patients (P = 0.004). In multivariate analysis, EGFR expression (P = 0.035) was found to be independent prognostic factors (significantly correlated to survival). CONCLUSION: EGFR overexpression was observed in 28.6% of Tunisian breast carcinoma, associated with unfavorable prognosis and with triple negative tumors. Systemically evaluation of EGFR in breast carcinoma could benefit especially to TN subgroup from EGFR targeting agents.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Receptores ErbB/metabolismo , Regulação Neoplásica da Expressão Gênica , Adulto , Idoso , Neoplasias da Mama/diagnóstico , Feminino , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica/métodos , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Receptor ErbB-2/biossíntese , Receptores de Estrogênio/biossíntese , Receptores de Progesterona/biossíntese , Resultado do Tratamento , Tunísia
19.
3 Biotech ; 12(3): 78, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35251881

RESUMO

Misidentification of human cell lines has previously led to confusing results during cell culture experiments. Although several enzymatic as well as molecular analysis approaches have been developed for cell-line authentication, these methods remain costly. In the present paper, we describe a simple chemical alternative based on known compound cell cytotoxicity. In addition to cisplatin, a pool of eight tamoxifen derivative compounds was used to compare the cytotoxic effects on three different breast cancer cell lines: MCF-7, T47D and MDA-MB-231. Our results show that four out of the eight cytotoxic-related compounds allowed to distinguish the different cell lines based on their IC50 (the half maximal inhibitory concentration) values which are cell type dependent. The remaining chemicals, particularly the most cytotoxic P15, showed close IC50 values for all the cell lines. Interestingly, flow cytometry experiments have identified notable differences among the three cell lines treated with P15. T47D and MDA-MB231 cells were blocked in SubG1 phase and S phase, respectively, while no significant change in cell cycle profile was noticed for MCF-7 cells. Differences were also noted at the level of caspase-3 activity and cell proliferation in P15-treated cells.

20.
Plant Physiol Biochem ; 189: 46-58, 2022 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-36044822

RESUMO

The nutritional enhancement of potato plants (Solanum tuberosum L.,) is highly critical. As it is considered a worldwide basic vegetarian nutrition to maintain health. S. tuberosum is one of the foremost staples and the world's fourth-largest food crop. In advance, its need is increasing because of its high-industrial value and population blast. To improve both potato growth and behavior under harsh environmental conditions, we produced transgenic potato plants overexpressing either VvNHX (a sodium proton antiporter from Vitis vinifera), VvCLC (a chloride channel from Vitis vinifera), or both. Control and transgenic plants were grown in greenhouse and field under non-stressed conditions for 85 days in order to characterize their phenotype and evaluate their agronomical performance. To this aim, the evaluation of plant growth parameters, tuber yields and characteristics (calibers, eye number and color), the chemical composition of tubers, was conducted and compared between the different lines. The obtained results showed that transgenic plants displayed an improved growth (flowering precocity, gain of vigor and better vegetative growth) along with enhanced tuber yields and quality (increased protein and starch contents). Our findings provide then insight into the role played by the VvNHX antiport and the VvCLC channel and a greater understanding of the effect of their overexpression in potato plants.


Assuntos
Solanum tuberosum , Antiporters/genética , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Canais de Cloreto/farmacologia , Tubérculos/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Trocadores de Sódio-Hidrogênio/metabolismo , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Amido/metabolismo
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