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1.
Adv Exp Med Biol ; 1131: 163-182, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31646510

RESUMO

Calcium (Ca2+) buffering is part of an integrative crosstalk between different mechanisms and elements involved in the control of free Ca2+ ions persistence in the cytoplasm and hence, in the Ca2+-dependence of many intracellular processes. Alterations of Ca2+ homeostasis and signaling from systemic to subcellular levels also play a pivotal role in the pathogenesis of many diseases.Compared with Ca2+ sequestration towards intracellular Ca2+ stores, Ca2+ buffering is a rapid process occurring in a subsecond scale. Any molecule (or binding site) with the ability to bind Ca2+ ions could be considered, at least in principle, as a buffer. However, the term Ca2+ buffer is applied only to a small subset of Ca2+ binding proteins containing acidic side-chain residues.Ca2+ buffering in the cytoplasm mainly relies on mobile and immobile or fixed buffers controlling the diffusion of free Ca2+ ions inside the cytosol both temporally and spatially. Mobility of buffers depends on their molecular weight, but other parameters as their concentration, affinity for Ca2+ or Ca2+ binding and dissociation kinetics next to their diffusional mobility also contribute to make Ca2+ signaling one of the most complex signaling activities of the cell.The crosstalk between all the elements involved in the intracellular Ca2+ dynamics is a process of extreme complexity due to the diversity of structural and molecular elements involved but permit a highly regulated spatiotemporal control of the signal mediated by Ca2+ ions. The basis of modeling tools to study Ca2+ dynamics are also presented.


Assuntos
Sinalização do Cálcio , Cálcio , Citoplasma , Animais , Tampões (Química) , Cálcio/metabolismo , Sinalização do Cálcio/fisiologia , Citoplasma/metabolismo , Humanos
2.
Postepy Biochem ; 65(3): 183-192, 2019 10 01.
Artigo em Polonês | MEDLINE | ID: mdl-31643165

RESUMO

The canonical Wnt pathway is related to regulation of embryogenesis, cell differentiation and proliferation. Various proteins are necessary for proper signal transduction and ß-catenin serves as the main mediator. In off-state of the Wnt pathway ß-catenin undergoes proteasomal degradation, while in on-state increase of cytoplasmic concentration of ß-catenin occurs followed by ß-catenin translocation into the cell nucleus. Interaction between ß-catenin and TCF/LEF transcription factors activates the expression of over hundred target genes of the Wnt pathway. Highly active Wnt signaling is observed in many cancers, including head and neck squamous cell carcinomas. Knowledge of the functional structure of the canonical Wnt pathway enables search of therapeutic molecular targets to effectively inhibit transcriptional activity of ß-catenin in cancer cells.


Assuntos
Via de Sinalização Wnt , beta Catenina/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Humanos , Neoplasias/metabolismo , Neoplasias/patologia , Fatores de Transcrição TCF/metabolismo , Via de Sinalização Wnt/genética
3.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 48(3): 318-325, 2019 May 25.
Artigo em Chinês | MEDLINE | ID: mdl-31496165

RESUMO

Proteins are the physical basis of life and perform all kinds of life activities. Proteins have different orientations and function in different tissues. The same protein, located in different subcellular regions, can perform different and even opposite functions. Both functional and structural proteins are capable of undergoing re-localization which can directly or indirectly participate in signal transduction. Due to abnormal transduction of signals during carcinogenesis, the proteins originally expressed in the cytoplasm are translocated into the nucleus and lead to functional changes in the tumor tissue. The changes of protein localization are affected by many factors, including the interaction between proteins, expression level of proteins and the cleaved intracellular domain of transmembrane protein.


Assuntos
Núcleo Celular , Citoplasma , Proteínas de Membrana , Carcinogênese/patologia , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Proteínas de Membrana/metabolismo , Domínios Proteicos , Transporte Proteico/fisiologia , Transdução de Sinais
4.
Chem Commun (Camb) ; 55(70): 10440-10443, 2019 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-31410422

RESUMO

In this work, a unique dual-site controlled fluorescent probe was presented for the sensitive and concurrent detection of pH in the cytoplasm and lysosomes. With the probe, the simultaneous down-regulation of pH in the lysosomes and cytoplasm during autophagy has been successfully revealed for the first time.


Assuntos
Autofagia , Citoplasma/metabolismo , Regulação para Baixo , Concentração de Íons de Hidrogênio , Lisossomos/metabolismo , Sondas Moleculares/química , Sobrevivência Celular , Células Hep G2 , Humanos , Espectrometria de Fluorescência
5.
Chemosphere ; 233: 786-795, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31340409

RESUMO

Microbial volatile organic compounds (mVCs) are formed in the metabolism of microorganisms and widely distributed in nature and pose threats to human health. However, the air pollution by microorganisms is a situation which is poorly understood. In this study, the cytotoxicity of E. aerogenes VCs was evaluated in the model organism Saccharomyces cerevisiae. E. aerogenes VCs inhibited the survival of yeast and triggered the formation of intracellular reactive oxygen species (ROS). The hypersensitive of MAP kinase mpk1/slt2 and 19S regulatory assembly chaperone adc17 mutants to the E. aerogenes VCs indicated cell wall integrity (CWI) pathway together with stress-inducible proteasome assembly regulation are essentially involved in mVCs tolerance mechanism. Furthermore, exposure to the mVCs resulted in the transcriptional upregulation of the CWI pathway, the regulatory particle assembly chaperones, and genes involved in proteasome regulations. Our research suggested that the ROS/MAPK signaling and proteasome regulatory pathway play pivotal roles in the integration and fine-tuning of the mVCs stress response. This study provides a molecular framework for future study of the effects of mVCs on more complex organisms, such as humans.


Assuntos
Enterobacter aerogenes/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Compostos Orgânicos Voláteis/farmacologia , Poluentes Atmosféricos/análise , Poluição do Ar/análise , Parede Celular/metabolismo , Citoplasma/metabolismo , Chaperonas Moleculares/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ativação Transcricional
6.
Cancer Invest ; 37(7): 293-298, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31328584

RESUMO

The aim of the study is to analyse the expression of Interleukin-6 receptor in different human gastric tissue and to correlate with the clinicopathological features of the patients. Immunohistochemistry was done against the IL-6R antibody and the Q-score was calculated from the staining pattern. Higher Q-scores were observed in tumour cells than the adjacent normal cells which were statistically significant. We also observed a significant correlation between the expressions of IL-6R and the clinicopathological features These findings suggest that IL-6R may represent as a therapeutic target for gastric carcinoma and serve as a prognostic indicator, as well.


Assuntos
Biomarcadores Tumorais/metabolismo , Receptores de Interleucina-6/metabolismo , Neoplasias Gástricas/metabolismo , Regulação para Cima , Adulto , Idoso , Idoso de 80 Anos ou mais , Membrana Celular/metabolismo , Citoplasma/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico
7.
Gene ; 710: 291-306, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31185283

RESUMO

The WD40 transcription factor family is a superfamily found in all eukaryotes that plays important roles in regulating growth and development. To our knowledge, to date, WD40 superfamily genes have been identified and characterized in several plant species, but little information is available on the WD40 superfamily genes in peach. In this study, we identified 220 members of the WD40 superfamily in the peach genome, and these members were further classified into five subfamilies based on phylogenetic comparison with those in Arabidopsis. The members within each subfamily had conserved motifs and gene structures. The WD40 genes were unevenly distributed on chromosomes 1 to 8 of the peach genome. Additionally, 58 pairs of paralog WD40 members were found on eight chromosomes in peach, and 242 pairs of orthologous WD40 genes in peach and Arabidopsis were matched. The 54 selected putative WD40 genes in peach had diverse expression patterns in red-fleshed and white-fleshed peach fruits at five developmental stages. Prupe.6G211800.1 was located only on the cytomembrane, while Prupe.1G428200.1 and Prupe.I003200.1 were located on both the cytomembrane and in the nucleus; Prupe.1G558700.1 was densely localized around the nuclear rim but relatively faintly localized in the nucleoplasm; Prupe.5G116300.1 was located in the nucleus and cytomembrane with strong signals but showed weak signals in the cytoplasm; and Prupe.8G212400.1 and Prupe.1G053600.1 were located mainly in the nuclear envelope and cytomembrane but relatively faintly in the nucleoplasm. This study provides a foundation for the further functional verification of WD40 genes in peach.


Assuntos
Mapeamento Cromossômico/métodos , Família Multigênica , Proteínas de Plantas/genética , Prunus persica/genética , Núcleo Celular/genética , Núcleo Celular/metabolismo , Cromossomos de Plantas/genética , Sequência Conservada , Citoplasma/genética , Citoplasma/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/metabolismo , Prunus persica/metabolismo , Repetições WD40
8.
Folia Histochem Cytobiol ; 57(2): 84-93, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31198984

RESUMO

INTRODUCTION: WNT5A (Wnt family member 5A) belongs to the WNT family of secreted signaling glycoproteins that play essential role in developmental, physiological and pathological processes. WNT5A was shown to take part in carcinogenesis process playing both oncogenic and suppressor functions in various types of human malignancies. This study aimed to assess the expression of the WNT5A gene at the mRNA and protein levels in the specimens derived from endometrial cancer (EC) or unchanged control endometrium. The associations between the WNT5A expression levels and clinicopathological characteristics and survival of EC patients were evaluated. MATERIALS AND METHODS: Total RNA was isolated in order to assess the relative amounts of WNT5A mRNA by quantitative polymerase chain reaction (QPCR) in samples of unchanged endometrial control (n = 8) and tumor samples of EC patients (n = 28). Immunohistochemistry (IHC) was used to determine the presence of WNT5A protein in the sections of formalin-fixed, paraffin-embedded tissue specimens derived from unchanged endome-trial controls (n = 6) and EC tumors (n = 19). Significance of differences in WNT5A expression levels between the studied groups of EC patients and correlations between the WNT5A and demographic data, pathological features, hematological parameters and overall survival of the patients were evaluated by statistical analysis. RESULTS: The level of WNT5A mRNA was decreased in EC in comparison to unchanged endometrium. WNT5A expression was associated with primary tumor invasion status exhibiting reduced level of transcripts in EC that involved organs beyond the uterus when compared to the uterus-confined cancers. WNT5A immunoreactivity was visualized in the cytoplasm and nuclei of EC cells as well as in the luminal and glandular epithelial cells of unchanged endometrium. WNT5A mRNA expression levels correlated negatively with cytoplasmic, and positively with nuclear immunoreactivity of the WNT5A protein in the EC cells. In addition, the relationships between blood leucocyte count (in particular granulocytes and lymphocytes) of patients with EC and their WNT5A mRNA and protein expression levels were established. A positive correlation between the nuclear immunoexpression of WNT5A protein in the cancer cells in cell nuclei and mean platelet volume in blood was also found. CONCLUSIONS: The results of the first study of WNT5A expression at the transcript and protein levels indicate that it could be considered as a potential marker of molecular changes that take place during EC development.


Assuntos
Neoplasias do Endométrio/genética , Neoplasias do Endométrio/metabolismo , RNA Mensageiro/metabolismo , Proteína Wnt-5a/genética , Proteína Wnt-5a/metabolismo , Adulto , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/imunologia , Biomarcadores Tumorais/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Regulação para Baixo , Neoplasias do Endométrio/diagnóstico , Neoplasias do Endométrio/mortalidade , Endométrio/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Prognóstico , Isoformas de Proteínas/genética , Isoformas de Proteínas/imunologia , Isoformas de Proteínas/metabolismo , Proteína Wnt-5a/imunologia
9.
Biochemistry (Mosc) ; 84(6): 583-592, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31238858

RESUMO

Actin plays an important role in cellular adhesion, muscle and non-muscle contractility, migration, polarization, mitosis, and meiosis. Investigation of specific mechanisms underlying these processes is essential not only for fundamental research but also for clinical applications, since modulations of actin isoforms are directly or indirectly correlate with severe pathologies. In this review we summarize the isoform-specific functions of actin associated with adhesion structures, motility and division of normal and tumor cells; alterations of the expression and structural organization of actin isoforms in normal and tumor cells. Selective regulation of cytoplasmic ß- or γ-actin expression determines functional diversity between isoforms: ß-actin plays the predominant role in contraction and intercellular adhesion, and γ-actin is responsible for the cellular plasticity and motility. Similar data were obtained in different epithelial and mesenchymal neoplastic cell cultures, as well as in immunomorphological comparison of normal human tissues with tumor analogues. Reorganization of the actin cytoskeleton and cell-cell contacts is essential for proliferation control and acquisition of invasiveness in epithelial tumors.


Assuntos
Actinas/fisiologia , Isoformas de Proteínas/fisiologia , Actinas/química , Animais , Adesão Celular/fisiologia , Movimento Celular/fisiologia , Transformação Celular Neoplásica , Citoplasma/metabolismo , Citosol/metabolismo , Humanos , Mamíferos , Isoformas de Proteínas/química , Relação Estrutura-Atividade
10.
Life Sci ; 231: 116543, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31176775

RESUMO

AIMS: CD155 is a ligand of the NK activating receptor DNAM-1, it has been described in a variety of human malignancies, but its expression in breast cancer remains unclear and poorly studied. MAIN METHODS: CD155 expression and NK cells infiltration were investigated in 158 patients with breast cancer by immunohistochemistry (IHC). Statistical analyses were performed to evaluate correlations of CD155 expression with clinical-pathological features, prognosis and tumor immunity. KEY FINDINGS: Tumor cytoplasmic CD155 (cyt-CD155) was associated with lymphovascular invasion (p = 0.011), and membranous CD155 (m-CD155) was strongly correlated with the presence of Tumor Infiltrating natural killer cells (NK-TILs) (p = 0.0003). Survival analysis demonstrated that patients with high cyt-CD155 had a significantly worse overall survival (p < 0.001) and death free survival (p = 0.014) than those with low expression, while high levels of m-CD155 correlated with a better prognosis (p = 0.037). Furthermore, we found that patients with m-CD155Low/NKLow tumors had a significantly reduced overall survival (p = 0.012). Multivariate analysis showed that positive tumor m-CD155 status was a significant independent marker of good prognosis. Meanwhile, high cyt-CD155 expression was identified as an independent poor prognostic predictor, suggesting a key role in this malignancy. SIGNIFICANCE: Altogether, our results revealed that cyt-CD155 was associated with invasiveness and poorer prognosis, but the concomitant presence of m-CD155 and NK-TILs had an opposite prognostic relevance in breast cancer. These results raised the importance of CD155 IHC analysis to elucidate biomarker localization, leading to better understand and design therapeutic molecule targeting CD155 in breast tumors.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Receptores Virais/genética , Receptores Virais/isolamento & purificação , Receptores Virais/metabolismo , Adulto , Idoso , Antígenos de Diferenciação de Linfócitos T/metabolismo , Neoplasias da Mama/imunologia , Citoplasma/metabolismo , Intervalo Livre de Doença , Feminino , Humanos , Imuno-Histoquímica , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Prognóstico , Receptores Virais/imunologia , Estudos Retrospectivos , Análise de Sobrevida
11.
Anticancer Res ; 39(6): 2805-2810, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31177117

RESUMO

BACKGROUND/AIM: Galectins belong to the family of galactose-binding proteins known to play an important role in the processes of cell proliferation, differentiation, migration and neoplastic progression. Herein, we studied the expression of galectin-3 (Gal-3) in chronic lymphocytic leukemia (CLL). MATERIALS AND METHODS: The expression of Gal-3 was analyzed by means of multiparametric flow cytometry in normal and pathological B-cells from peripheral blood and bone marrow samples of 67 patients with CLL. RESULTS: Pathological B-cells expressed significantly higher levels of cytoplasmic Gal-3 than normal B-cells. Moreover, overexpression of cytoplasmic Gal-3 was observed in the prognostically poorest subgroup of CLL patients, namely those with 17p deletion. CONCLUSION: Our results indicate a possible role of galectin-3 in CLL pathophysiology and its potential value as a prognostic marker and therapeutic target.


Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 17/genética , Galectina 3/genética , Leucemia Linfocítica Crônica de Células B/genética , Regulação para Cima , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Citoplasma/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico
12.
Anticancer Res ; 39(6): 2811-2819, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31177118

RESUMO

BACKGROUND/AIM: Recent knowledge implicates a differential expression of the insulin-like growth factor-I (IGF-I) mRNA splice variants (i.e., IGF-IEa, IGF-IEb and IGF-IEc) in cancerous tissues, implying possible specific roles of the encoded IGF-I protein isoforms in cancer biology. In particular, there is growing evidence that the IGF-IEc isoform may play a distinct biological role in various types of cancers. The present study investigated whether IGF-IEc expression is associated with a particular type of thyroid cancer. MATERIALS AND METHODS: Formalin-fixed paraffin-embedded tissue specimens of different types of thyroid cancers from 92 patients were assessed for IGF-IEc expression by immunohistochemistry. In addition, thyroid cancer biopsies of different TNM staging histological types were evaluated for mRNA expression of the IGF-IEc transcript by real-time polymerase chain reaction (PCR). RESULTS: From the total number of 92 samples, 2 were anaplastic, 10 medullary, 4 hyperplasias of C-cells, 11 follicular, 5 hurtle cell carcinomas, 2 poorly differentiated, 5 nodular hyperplasias, 1 lymphoma and 52 were papillary thyroid cancers. The age of cancer diagnosis or tumor size did not significantly affect the IGF-IEc expression. Among all types of cancers, IGF-IEc was expressed in papillary differentiated thyroid cancer. Its expression/localization was mainly cytoplasmic and significantly associated with TNM staging and the presence of muscular and capsule cancerous invasion (p<0.05). Similarly, a differential profile was revealed regarding the mRNA expression of the IGF-IEc transcript, that exhibited a higher expression in aggressive compared to the non-aggressive papillary cancers. CONCLUSION: IGF-IEc isoform expression in thyroid cancer is positively associated with more advanced stages of papillary thyroid cancer.


Assuntos
Processamento Alternativo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Câncer Papilífero da Tireoide/patologia , Neoplasias da Glândula Tireoide/patologia , Regulação para Cima , Adolescente , Adulto , Idoso , Citoplasma/genética , Citoplasma/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Retrospectivos , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/metabolismo , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/metabolismo , Carga Tumoral , Adulto Jovem
13.
Plant Physiol Biochem ; 141: 206-214, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31176880

RESUMO

SUMOylation is an important protein modification that regulates the properties of substrate proteins in a variety of cellular processes. SUMOylation is catalyzed via a cascade of enzymes and is usually stimulated by SUMO E3 ligases. However, the molecular functions and regulatory mechanisms of SUMOylation in forage crops are unknown. Here, we isolated and functionally characterized DiMMS21, a homolog of the Arabidopsis thaliana SUMO ligase AtMMS21, from the forage legume Desmodium intortum. DiMMS21 is expressed ubiquitously in various D. intortum organs and its encoded protein is found in the cytoplasm and nucleus. Bioinformatics analysis indicated that DiMMS21 contains a conserved SP-RING domain that is required for its activity. Biochemical evidence supports the notion that this protein is a functional SUMO ligase. When expressed in an Arabidopsis mms21 mutant, DiMMS21 completely rescued the defects in root, leaf, and silique development. The results from cotyledon greening and marker gene expression suggested that DiMMS21 can only partially complements the role of AtMMS21 in abscisic acid (ABA) responses. In summary, we characterized the molecular features of DiMMS21 and uncovered potential roles of this SUMO ligase in development and ABA responses, increasing our understanding on the function of SUMOylation in forage crops.


Assuntos
Fabaceae/enzimologia , Regulação da Expressão Gênica de Plantas , Ligases/metabolismo , Proteínas de Plantas/metabolismo , Ácido Abscísico/metabolismo , Arabidopsis/enzimologia , Proteínas de Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Biologia Computacional , Citoplasma/metabolismo , DNA Complementar/metabolismo , Perfilação da Expressão Gênica , Mutação , Filogenia , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Sementes/metabolismo , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo
14.
Biochemistry (Mosc) ; 84(6): 593-607, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31238859

RESUMO

Mitochondria are among the most important cell organelles involved in the regulation of intracellular calcium homeostasis. During the last decade, a number of molecular structures responsible for the mitochondrial calcium transport have been identified including the mitochondrial Ca2+ uniporter (MCU), Na+/Ca2+ exchanger (NCLX), and Ca2+/H+ antiporter (Letm1). The review summarizes the data on the structure, regulation, and physiological role of such structures. The pathophysiological mechanism of Ca2+ transport through the cyclosporine A-sensitive mitochondrial permeability transition pore is discussed. An alternative mechanism for the mitochondrial pore opening, namely, formation of the lipid pore induced by saturated fatty acids, and its role in Ca2+ transport are described in detail.


Assuntos
Cálcio/metabolismo , Mitocôndrias/metabolismo , Animais , Canais de Cálcio/metabolismo , Citoplasma/metabolismo , Humanos , Transporte de Íons , Metabolismo dos Lipídeos , Membranas Mitocondriais/metabolismo , Estrutura Molecular , Trocador de Sódio e Cálcio/metabolismo
15.
Chem Commun (Camb) ; 55(54): 7852-7855, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31215553
16.
Nanomedicine (Lond) ; 14(10): 1247-1265, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31084385

RESUMO

Aim: Plant virus-like particles (VLPs) have emerged as a novel platform for delivery of drugs/antibodies. The aim of the present investigation is to establish the entry mechanism of flexuous rod-shaped virus particles into mammalian cells. Methods: Far-Western blot analysis, pull-down and ELISA were used to characterize vimentin and Hsp60 interaction with VLPs. The mode/kinetics of internalization of VLPs was deciphered using pharmacological inhibitors/endosomal markers. Results & discussion: The flexuous rod-shaped VLPs of Pepper vein banding virus (PVBV) enter HeLa and HepG2 cells via cell-surface proteins: vimentin and Hsp60, respectively. VLPs internalize via different modes of endocytosis in HeLa, HepG2 cells and are biodegradable. Vimentin and Hsp60 could be potential epithelial ligands that facilitate targeting of nanoparticles to tumor cells.


Assuntos
Endocitose , Células Epiteliais/metabolismo , Nanopartículas/metabolismo , Potyvirus/fisiologia , Animais , Transporte Biológico , Chaperonina 60/metabolismo , Citoplasma/metabolismo , Células HeLa , Células Hep G2 , Humanos , Cinética , Vimentina/metabolismo , Vírion/fisiologia , Internalização do Vírus
17.
BMC Plant Biol ; 19(1): 175, 2019 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-31046676

RESUMO

BACKGROUND: DUOII is a multi-ovary wheat (Triticum aestivum L.) line with two or three pistils and three stamens in each floret. The multi-ovary trait of DUOII is controlled by a dominant gene, whose expression can be suppressed by the heterogeneous cytoplasm of TeZhiI (TZI), a line with the nucleus of common wheat and the cytoplasm of Aegilops. Crosses between female DUOII plants and male TZI plants resulted in multi-ovary F1s; whereas, the reciprocal crosses resulted in mono-ovary F1s. Although the multi-ovary trait is inherited as single trait controlled by a dominant allele in lines with a Triticum cytoplasm, the mechanism by which the special heterogeneous cytoplasm suppresses the expression of multi-ovary is not well understood. RESULTS: Observing the developmental process, we found that the critical stage of additional pistil primordium development was when the young spikes were 2-6 mm long. Then, we compared the quantitative proteomic profiles of 2-6 mm long young spikes obtained from the reciprocal crosses between DUOII and TZI. A total of 90 differentially expressed proteins were identified and analyzed based on their biological functions. These proteins had obvious functional pathways mainly implicated in chloroplast metabolism, nuclear and cell division, plant respiration, protein metabolism, and flower development. Importantly, we identified two key proteins, Flowering Locus K Homology Domain and PEPPER, which are known to play an essential role in the specification of pistil organ identity. By drawing relationships between the 90 differentially expressed proteins, we found that these proteins revealed a complex network which is associated with multi-ovary gene expression under heterogeneous cytoplasmic suppression. CONCLUSIONS: Our proteomic analysis has identified certain differentially expressed proteins in 2-6 mm long young spikes, which was the critical stage of additional primordium development. This paper provided a universal proteomic profiling involved in the cytoplasmic suppression of wheat floral meristems; and our findings have laid a solid foundation for further mechanistic studies on the underlying mechanisms that control the heterogeneous cytoplasm-induced suppression of the nuclear multi-ovary gene in wheat.


Assuntos
Citoplasma/metabolismo , Triticum/metabolismo , Cruzamentos Genéticos , Flores/anatomia & histologia , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Proteômica , Triticum/anatomia & histologia , Triticum/genética
18.
Med Oncol ; 36(7): 61, 2019 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-31140031

RESUMO

Testicular germ cell tumors are the most frequent malignancies found in men between 15 and 44 years old. Although cellular apoptosis susceptibility (CAS) was demonstrated to be upregulated in breast cancer and colon cancer, the expression of CAS in the human testis and testicular germ cell tumors remained elusive. In the present study, CAS-positive signals were detected in the normal testicular tissues, cancer adjacent normal testicular tissues, seminoma, yolk sac tumor, and teratoma. Interestingly, the expression level of CAS in testicular germ cell tumors (TGCTs) (but not seminoma) was significantly lower than that of human testicular tissues and cancer adjacent normal testicular tissues, suggesting that decreased CAS contributed to the progression of TGCTs. Notably, the expression of CAS in seminoma was significantly higher than that of in the non-seminomas, consistent with the results from TCGA database. Furthermore, the localization of CAS is mainly restricted in the nucleus in the lesions of normal human testicular tissue and cancer adjacent normal testicular tissue. Although the expression of CAS was not significantly different between normal testicular tissue and seminoma, CAS was more enriched in cytoplasm in seminoma compared to the normal, cancer adjacent tissue and other types of TGCTs. The current results demonstrated reduced expression of CAS in the human testicular germ cell tumors and the CAS translocation from the nuclear to cytoplasm in seminoma, thereby supporting a possible role in normal testis function and in the development of seminoma.


Assuntos
Proteína de Suscetibilidade a Apoptose Celular/biossíntese , Neoplasias Embrionárias de Células Germinativas/metabolismo , Neoplasias Testiculares/metabolismo , Testículo/metabolismo , Carcinoma Embrionário/metabolismo , Citoplasma/metabolismo , Tumor do Seio Endodérmico/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Seminoma/metabolismo , Teratoma/metabolismo , Análise Serial de Tecidos
19.
Molecules ; 24(9)2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31071981

RESUMO

Atherosclerosis is the pathological basis of cardiovascular disease, whilst endothelial dysfunction (ED) plays a primary role in the occurrence and development of atherosclerosis. Simvastatin has been shown to possess significant anti-atherosclerosis activity. In this study, we evaluated the protective effect of simvastatin on endothelial cells under oxidative stress and elucidated its underlying mechanisms. Simvastatin was found to attenuate H2O2-induced human umbilical vein endothelial cells (HUVECs) dysfunction and inhibit the Wnt/ß-catenin pathway; however, when this pathway was activated by lithium chloride, endothelial dysfunction was clearly enhanced. Further investigation revealed that simvastatin did not alter the expression or phosphorylation of LRP6, but reduced intracellular cholesterol deposition and inhibited endoplasmic reticulum (ER) stress. Inducing ER stress with tunicamycin activated the Wnt/ß-catenin pathway, whereas reducing ER stress with 4-phenylbutyric acid inhibited it. We hypothesize that simvastatin does not affect transmembrane signal transduction in the Wnt/ß-catenin pathway, but inhibits ER stress by reducing intracellular cholesterol accumulation, which blocks intracellular signal transduction in the Wnt/ß-catenin pathway and ameliorates endothelial dysfunction.


Assuntos
Estresse do Retículo Endoplasmático/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/patologia , Peróxido de Hidrogênio/toxicidade , Sinvastatina/farmacologia , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colesterol/metabolismo , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Modelos Biológicos , Estresse Oxidativo/efeitos dos fármacos , Via de Sinalização Wnt/efeitos dos fármacos
20.
Saudi Med J ; 40(5): 432-439, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31056618

RESUMO

OBJECTIVES:   To evaluate p63 expression pattern in Saudi colorectal cancer (CRC) patients and correlate that with clinicopathological parameters and its role in carcinogenesis and prognosis. METHODS: Archival tumor samples were analyzed by immunohistochemistry for p63 expression in 324 consecutive Saudi patients diagnosed with CRC between January 2006 and December 2017 at the Pathology Department of a tertiary care Hospital, Madinah, Saudi Arabia. RESULTS: P63 over-expression was absent in normal mucosa, while 12.5% cases of adenoma showed its over-expression. In CRC, p63 expression was high in 24.1% of cases. There were no significant correlations between p63 expression and gender, tumor location, tumor size and tumor histologic differentiation. However, high p63 expression revealed a significant correlation with age (p=0.035), tumor type (p=0.004), American Joint Committee on Cancer stage (p=0.046), lymph node metastasis (p=0.006), lymphovascular invasion (p=0.006), distant metastasis (p=0.049), high Ki67 expression (p=0.000) and K-ras expression (p=0.002). The Kaplan-Meier analysis revealed a shorter period of survival with p63 over-expression (p less than 0.001). The Cox-regression model analysis showed that p63 over-expression was an independent prognostic marker in CRC (p=0.000). CONCLUSION: P63 expression was increased from normal to adenoma to carcinoma sequence. Moreover, p63 cytoplasmic expression seems to be related to high Ki67 indexing, K-ras expression, advanced tumor stage and poor clinical outcome of CRC. These findings suggest a significant role of cytoplasmic p63 expression in tumor progression and prognosis.


Assuntos
Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/genética , Citoplasma/genética , Citoplasma/metabolismo , Expressão Gênica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/patologia , Feminino , Humanos , Imuno-Histoquímica , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Prognóstico , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Taxa de Sobrevida , Adulto Jovem
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