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1.
Mol Biol Evol ; 40(4)2023 04 04.
Artículo en Inglés | MEDLINE | ID: mdl-36929909

RESUMEN

Photic niche shifts of mammals are associated with changing visual capabilities, primarily mediated by three visual pigments, two (SWS1 and M/LWS) of them for color vision and rhodopsin (RH1) for dim-light vision. To further elucidate molecular mechanisms of mammalian visual adaptations to different light environments, a systematic study incorporating evolutionary analyses across diverse groups and in vitro assays have been carried out. Here, we collected gene sequences for the three opsins from 220 species covering all major mammalian clades. After screening for cone opsin gene losses, we estimated selective pressures on each of the three genes and compared the levels of selection experienced by species living in bright- and dim-light environments. SWS1 pigment is shown to experience accelerated evolution in species living in bright-light environments as has RH1 in aquatic cetaceans, indicating potential shifts for ecological adaptations. To further elucidate the functional mechanisms for these two pigments, we then carried out site-directed mutagenesis in representative taxa. For SWS1, violet and ultraviolet sensitivities in the pika and mouse are mainly affected by substitutions at the critical sites 86 and 93, which have strong epistatic interaction. For RH1, the phenotypic difference between the sperm whale and bovine sequences is largely contributed by a substitution at site 195, which could be critical for dim-light sensation for deep-diving species. Different evolutionary patterns for the visual pigments have been identified in mammals, which correspond to photic niches, although additional phenotypic assays are still required to fully explain the functional mechanisms.


Asunto(s)
Evolución Molecular , Mamíferos , Animales , Bovinos , Ratones , Filogenia , Opsinas/genética , Rodopsina/genética
2.
Bioorg Chem ; 141: 106836, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37774436

RESUMEN

Alzheimer's diseases (AD) and other infectious diseases caused by drug-resistance bacteria have posed a serious threat to human lives and global health. With the aim to search for human acetylcholinesterase (hAChE) inhibitors and antibacterial agents from medicinal plants, 16 phloroglucinol oligomers, including two new phloroglucinol monomers (1a and 1b), four new phloroglucinol dimers (3a, 3b, 4b, and 5a), six new phloroglucinol trimers (6a, 6b, 7a, 7b, 8a, and 8b), and two naturally occurring phloroglucinol monomers (2a and 2b), along with two known congeners (4a and 5b), were purified from the leaves of tropic Rhodomyrtus tomentosa. The structures and absolute configurations of these new isolates were unequivocally established by comprehensive analyses of their spectroscopic data (NMR and HRESIMS), ECD calculation, and single crystal X-ray diffraction. Structurally, 3a/3b shared a rare C-5' formyl group, whereas 6a/6b possessed a unique C-7' aromatic ring. In addition, 7a/7b and 8a/8b were rare phloroglucinol trimers with a bis-furan and a C-6' hemiketal group. Pharmacologically, the mixture of 3a and 3b showed the most potent human acetylcholinesterase (hAChE) inhibitory activity with an IC50 value of 1.21 ± 0.16 µM. The molecular docking studies of 3a and 3b in the hAChE binding sites were performed, displaying good agreement with the in vitro inhibitory effects. In addition, the mixture of 3a and 3b displayed the most significant anti-MRSA (methicillin-resistant Staphylococcus aureus) with MIC and MBC values of both 0.50 µg/mL, and scanning electron microscope (SEM) studies revealed that they could destroy the biofilm structures of MRSA. The findings provide potential candidates for the further development of anti-AD and anti-bacterial agents.


Asunto(s)
Antibacterianos , Inhibidores de la Colinesterasa , Staphylococcus aureus Resistente a Meticilina , Floroglucinol , Humanos , Acetilcolinesterasa , Antibacterianos/farmacología , Simulación del Acoplamiento Molecular , Estructura Molecular , Floroglucinol/análogos & derivados , Floroglucinol/química , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/farmacología , Extractos Vegetales/química
3.
World J Surg Oncol ; 21(1): 203, 2023 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-37430268

RESUMEN

PURPOSE: Thymoma is the most common primary tumor in the anterior mediastinum. The prognostic factors of patients with thymoma still need to be clarified. In this study, we aimed to investigate the prognostic factors of patients with thymoma who received radical resection and establish the nomogram to predict the prognosis of these patients. MATERIALS AND METHODS: Patients who underwent radical resection for thymoma with complete follow-up data between 2005 and 2021 were enrolled. Their clinicopathological characteristics and treatment methods were retrospectively analyzed. Progression-free survival (PFS) and overall survival (OS) were estimated using the Kaplan-Meier method and compared by the log-rank test. Univariate and multivariate Cox proportional hazards regression analyses were performed to identify the independent prognostic factors. According to the results of the univariate analysis in the Cox regression model, the predictive nomograms were created. RESULTS: A total of 137 patients with thymoma were enrolled. With a median follow-up of 52 months, the 5-year and 10-year PFS rates were 79.5% and 68.1%, respectively. The 5-year and 10-year OS rates were 88.4% and 73.1%, respectively. Smoking status (P = 0.022) and tumor size (P = 0.039) were identified as independent prognostic factors for PFS. Multivariate analysis showed that a high level of neutrophils (P = 0.040) was independently associated with OS. The nomogram showed that the World Health Organization (WHO) histological classification contributed more to the risk of recurrence than other factors. Neutrophil count was the most important predictor of OS in patients with thymoma. CONCLUSION: Smoking status and tumor size are risk factors for PFS in patients with thymoma. A high level of neutrophils is an independent prognostic factor for OS. The nomograms developed in this study accurately predict PFS and OS rates at 5 and 10 years in patients with thymoma based on individual characteristics.


Asunto(s)
Timoma , Neoplasias del Timo , Humanos , Timoma/cirugía , Pronóstico , Estudios Retrospectivos , Neoplasias del Timo/cirugía , Organización Mundial de la Salud
4.
Zhongguo Zhong Yao Za Zhi ; 47(13): 3519-3525, 2022 Jul.
Artículo en Zh | MEDLINE | ID: mdl-35850804

RESUMEN

The present study investigated the chemical constituents and inhibitory activities against α-glucosidase from the seeds of Morinda citrifolia(Noni) by the chromatographic technique and semi-preparative HPLC.Fifteen compounds were isolated from the ethyl acetate extract of the seeds, and their structures were identified on the basis of physiochemical characteristics and spectroscopic data as(9S,2E,4Z,7Z)-9-hydroxydeca-2,4,7-trienoic acid(1), azelaic acid(2), scopoletin(3), ursolic acid(4), quercetin(5), cyclo-(L-Leu-L-Ile)(6), cyclo-(L-Phe-L-Ile)(7), cyclo-(L-Phe-L-Val)(8), cyclo-(L-Leu-L-Val)(9), cyclo-(L-Phe-L-Leu)(10), caffeic acid(11), 3,4-dihydroxycinnamaldehyde(12), p-hydroxybenzoic acid(13), p-hydroxy-cinnamic acid(14), and p-hydroxyphenethyl alcohol(15).Among them, compound 1 was a new fatty acid and compounds 7-10 and 12 were isolated from Morinda plant in the Rubiaceae family for the first time.Compounds 1, 2 and 4-15 were isolated from the seeds of M.citrifolia(Noni) for the first time.All isolated compounds were evaluated for the inhibitory activities against α-glucosidase and compounds 3-5 showed potential inhibitory activity with IC_(50) values of 160, 133, and 120 µmol·L~(-1), respectively.


Asunto(s)
Morinda , Frutas/química , Morinda/química , Extractos Vegetales/química , Escopoletina , Semillas/química , alfa-Glucosidasas
5.
J Mol Evol ; 89(7): 458-471, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34287664

RESUMEN

Mammals have evolved different tooth phenotypes that are hypothesized to be associated with feeding habits. However, the genetic basis for the linkage has not been well explored. In this study, we investigated 13 tooth-related genes, including seven enamel-related genes (AMELX, AMBN, ENAM, AMTN, ODAM, KLK4 and MMP20) and six dentin-related genes (DSPP, COL1A1, DMP1, IBSP, MEPE and SPP1), from 63 mammals to determine their evolutionary history. Our results showed that different evolutionary histories have evolved among divergent feeding habits in mammals. There was stronger positive selection for eight genes (ENAM, AMTN, ODAM, KLK4, DSPP, DMP1, COL1A1, MEPE) in herbivore lineages. In addition, AMELX, AMBN, ENAM, AMTN, MMP20 and COL1A1 underwent accelerated evolution in herbivores. While relatively strong positive selection was detected in IBSP, SPP1, and DSPP, accelerated evolution was only detected for MEPE and SPP1 genes among the carnivorous lineages. We found positive selection on AMBN and ENAM genes for omnivorous primates in the catarrhini clade. Interestingly, a significantly positive association between the evolutionary rate of ENAM, ODAM, KLK4, MMP20 and the average enamel thickness was found in primates. Additionally, we found molecular convergence in some amino acid sites of tooth-related genes among the lineages whose feeding habit are similar. The positive selection of related genes might promote the formation and bio-mineralization of tooth enamel and dentin, which would make the tooth structure stronger. Our results revealed that mammalian tooth-related genes have experienced variable evolutionary histories, which provide some new insights into the molecular basis of dietary adaptation in mammals.


Asunto(s)
Proteínas del Esmalte Dental , Evolución Molecular , Animales , Mamíferos/genética , Fenotipo , Primates
6.
Bioorg Chem ; 117: 105404, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34749116

RESUMEN

Alzheimer's disease (AD) diagnoses are greatly increasing in frequency as the global population ages, highlighting an urgent need for new anti-AD strategies. With the aim to search for human acetylcholinesterase (hAChE) inhibitors from the species of Myrtaceae family, ten acylphloroglucinol trimers (APTs), including eight new APTs, callistemontrimers A-H (1a, 1b, 2a, 2b, 3a, 3b, 4b, and 5b), and two naturally occurring ones (4a and 5a), along with one reported triketone-acylphloroglucinol-monoterpene adduct (6), were obtained and structurally characterized from the hAChE inhibitory acetone extract of Callistemon salignus seeds. The structures and their absolute configurations for new APTs were unequivocally established via the detailed interpretation of extensive spectroscopic data (HRESIMS and NMR), ECD calculations, and single crystal X-ray diffraction, whereas the absolute configurations of known APTs were determined by further chiral separation, and calculated ECD calculations. The results of hAChE inhibitory assay revealed that an enantiomeric mixture of 2a/2b, 2a, and 2b are good hAChE inhibitors with IC50 values of 1.22 ±â€¯0.23, 2.28 ±â€¯0.19, and 4.96 ±â€¯0.39 µM, respectively. Molecular docking was used to uncover the modes of interactions for bioactive compounds with the active site of hAChE. In addition, 2 and 6 displayed moderate neurite outgrowth-promoting effects with differentiation rates of 6.16% and 6.19% at a concentration of 1.0 µM, respectively.


Asunto(s)
Inhibidores de la Colinesterasa/farmacología , Floroglucinol/farmacología , Acetilcolinesterasa/metabolismo , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/aislamiento & purificación , Humanos , Simulación del Acoplamiento Molecular , Myrtaceae/química , Floroglucinol/análogos & derivados , Floroglucinol/aislamiento & purificación , Estereoisomerismo , Relación Estructura-Actividad
7.
J Nat Prod ; 79(6): 1628-34, 2016 06 24.
Artículo en Inglés | MEDLINE | ID: mdl-27203291

RESUMEN

Twelve new diterpenoids based on two rare skeletal types, namely, paralianones A-D (1-4) and pepluanols A-H (5-12), along with five known compounds, were isolated from an acetone extract of Euphorbia peplus. Their structures were proposed based on 1D and 2D NMR spectroscopic data analysis. These diterpenoids were evaluated for potential anti-inflammatory activity in a lipopolysaccharide-stimulated mouse macrophage cellular model. Compounds 3, 4, 11, 13, and 16 displayed moderate inhibitory effects on NO inhibition, with IC50 values ranging from 29.9 to 38.3 µM.


Asunto(s)
Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Diterpenos/aislamiento & purificación , Diterpenos/farmacología , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Euphorbia/química , Animales , Antiinflamatorios/química , Antineoplásicos Fitogénicos/química , Diterpenos/química , Medicamentos Herbarios Chinos/química , Concentración 50 Inhibidora , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Ratones , Estructura Molecular , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico/biosíntesis , Raíces de Plantas/química
8.
Molecules ; 21(9)2016 Aug 26.
Artículo en Inglés | MEDLINE | ID: mdl-27571056

RESUMEN

Three new triterpenoid alkaloids, namely buxmicrophyllines P-R (1-3), were isolated from the twigs and leaves of Buxus microphylla. Their structures were elucidated on the basis of NMR and MS spectroscopic analyses. Structurally, compounds 1-3 belong to the 9,10-cycloartane type alkaloids. In addition, compound 3 exhibited moderate cytotoxic activities in vitro against HL-60, SMMC-7221, A-549, MCF-7, and SW480 cell lines (with IC50 values ranging from 4.51 to 15.58 µM).


Asunto(s)
Alcaloides , Antineoplásicos Fitogénicos , Buxus/química , Citotoxinas , Triterpenos , Células A549 , Alcaloides/química , Alcaloides/aislamiento & purificación , Alcaloides/farmacología , Antineoplásicos Fitogénicos/química , Antineoplásicos Fitogénicos/aislamiento & purificación , Antineoplásicos Fitogénicos/farmacología , Citotoxinas/química , Citotoxinas/aislamiento & purificación , Citotoxinas/farmacología , Células HL-60 , Humanos , Células MCF-7 , Triterpenos/química , Triterpenos/aislamiento & purificación , Triterpenos/farmacología
9.
Yi Chuan ; 38(5): 418-26, 2016 05.
Artículo en Zh | MEDLINE | ID: mdl-27232490

RESUMEN

Phytohormones are signaling molecules that control plant growth and development. Recent studies revealed that non-coding small RNAs play critical roles in plant development and stress responses via phytohormone signaling pathways. In this review, we summarize the present knowledge on the microRNAs (miRNAs) and secondary short interfering RNAs (siRNAs) involved in phytohormone signaling pathways, which include auxin, gibberellic acid, brassinosteroid and abscisic acid pathways. We also discuss their possible implications in phytohormone crosstalk during specific developmental processes.


Asunto(s)
MicroARNs/fisiología , Reguladores del Crecimiento de las Plantas/fisiología , ARN Interferente Pequeño/fisiología , Transducción de Señal/fisiología , Animales , Humanos , Desarrollo de la Planta
10.
Yi Chuan ; 38(8): 756-64, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27531614

RESUMEN

IAA2 is a member of the Aux/IAA auxin responsive gene family in Arabidopsis thaliana. No iaa2 mutant has been reported until now, thus hindering its further mechanistic investigations. The normal genomic editing technology of CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) uses only a single guide RNA (sgRNA) to target one site in a specific gene, and the gene knockout efficiency is not high. Instead, multiple sgRNAs can target multiple sites; therefore, the efficiency may be improved. In the present investigation, we used the golden-gate cloning strategy and two rounds of PCR reactions to combine three sgRNAs in the same entry vector. The final expression vector was obtained by LR reactions with the destination vector containing the Cas9 expression cassette. Four out of the six sgRNAs were effective, and we also obtained a lot of insertion and deletion mutations. Compared with one sgRNA approach, multiple sgRNAs displayed higher gene-knockout efficiency and produced more germ-line mutants. Thus, we established a more rapid and efficient method and generated five mutants for further studies of IAA2 functions.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Vectores Genéticos/genética , Edición de ARN/genética , ARN Guía de Kinetoplastida/genética , Factores de Transcripción/genética , Secuencia de Bases , Sistemas CRISPR-Cas/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , Edición Génica/métodos , Técnicas de Inactivación de Genes/métodos , Mutación/genética
11.
Yi Chuan ; 38(7): 644-650, 2016 07 20.
Artículo en Inglés | MEDLINE | ID: mdl-27733337

RESUMEN

Autophagy is an evolutionarily highly conserved catabolic pathway among eukaryotic cells that protects the organisms against environmental stress. Normally, autophagy is mainly involved with autophagy-related proteins(ATGs) and autophagic regulators including a series of cytoplasmic proteins and small molecules. Besides, the selective autophagy, which targets damaged organalles or protein aggregates, is mediated by the additional receptors to help the ATGs recognize different substrates. In this review, we summarize recent advances in autophagic regulators like ROS(Reactive oxygen species), TOR(Target of rapamycin) and receptors like NBR1(Neighbor of BRCA1 gene protein), RPN10(Regulatory particle non-ATPase 10) as well as their functional mechanisms mainly in Arabidopsis thaliana.


Asunto(s)
Autofagia , Proteínas de Plantas/fisiología , Proteínas de Arabidopsis/fisiología , Proteínas Portadoras/fisiología , Humanos , Péptidos y Proteínas de Señalización Intracelular , Proteínas/fisiología , Serina-Treonina Quinasas TOR/fisiología , Proteínas de Transporte Vesicular/fisiología
12.
Infect Drug Resist ; 17: 1171-1184, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38544964

RESUMEN

Background: The surge in the number of patients diagnosed with COVID-19 since China's open-door policy has placed a huge burden on the public healthcare system, especially the intensive care system. This study's objective was to discover possible clinical outcome predictors in COVID-19 patients treated in intensive care units (ICUs) and to provide useful information for future preventative efforts and therapies. Methods: This retrospective study included 173 COVID-19 critically ill patients and reviewed the 28-day survival outcome in the First Affiliated Hospital of Nanjing Medical University. Competing risk analysis was performed to predict the cumulative incidence function (CIF) of mortality in hospital. The independent prognostic factors were identified by applying the Fine-Gray proportional subdistribution hazard model. Receiver operating characteristic (ROC) curves were used to evaluate model efficacy, and calibration curves were used to validate the model. Finally, we compared the competing risk model with the traditional proportional hazards model (Cox regression model) using CIF. Results: Of these 173 patients, 66 (38.2%) survived, 55 (31.8%) died, and 52 (30.0%) discharged. In univariate analysis, 12 variables were significantly correlated with mortality. In multivariate analysis, Age, Neutrophil ratio, Direct Bilirubin (DBIL) and Renal disease were independent predictors of 28-day outcome. The ROC curve of the multivariate prediction model showed an AUC (area under the curve) of 0.790. The results of the calibration curve and the concordance index (C-index) show that the model has good discriminatory power. The competing risk model we applied was more accurate than the Cox model. Conclusion: We presented a more accurate multivariate prediction model for 28-day in-hospital mortality for ICU COVID-19 patients using a competing risk model.

13.
Front Genet ; 15: 1315677, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38725483

RESUMEN

To cope with the damage from oxidative stress caused by hypoxia, mammals have evolved a series of physiological and biochemical traits, including antioxidant ability. Although numerous research studies about the mechanisms of hypoxia evolution have been reported, the molecular mechanisms of antioxidase-related genes in mammals living in different environments are yet to be completely understood. In this study, we constructed a dataset comprising 7 antioxidase-related genes (CAT, SOD1, SOD2, SOD3, GPX1, GPX2, and GPX3) from 43 mammalian species to implement evolutionary analysis. The results showed that six genes (CAT, SOD1, SOD2, SOD3, GPX1, and GPX3) have undergone divergent evolution based on the free-ratio (M1) model. Furthermore, multi-ratio model analyses uncovered the divergent evolution between hypoxic and non-hypoxic lineages, as well as various hypoxic lineages. In addition, the branch-site model identified 9 positively selected branches in 6 genes (CAT, SOD1, SOD2, SOD3, GPX2, and GPX3) that contained 35 positively selected sites, among which 31 positively selected sites were identified in hypoxia-tolerant branches, accounting for 89% of the total number of positively selected sites. Interestingly, 65 parallel/convergent sites were identified in the 7 genes. In summary, antioxidase-related genes are subjected to different selective pressures among hypoxia-tolerant species living in different habitats. This study provides a valuable insight into the molecular evolution of antioxidase-related genes in hypoxia evolution in mammals.

14.
Biochem Cell Biol ; 91(6): 435-42, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24219285

RESUMEN

Angiotensin converting enzyme II (ACE2) is a component of the renin-angiotensin system (RAS) that negatively regulates angiotensin II (Ang II). Ang II, in turn, affects the expression of matrix metalloproteinases (MMPs) to induce heart remodeling. The specific mechanisms by which ACE2 regulates MMP-2, however, remain unclear. The aim of this study was to investigate the regulatory relationships between Ang II, ACE2, and MMP-2. ACE2 expression was upregulated and downregulated in human cardiofibroblasts (HCFs) by lentiviral infection. Effects on MMP-2 activity, shed ACE2 activity, extracellular signal-regulated kinase (ERK) signaling pathway, and ADAM metallopeptidase domain 17 (ADAM17) expression were assessed. ACE2 increased MMP-2 activity, and Ang II inhibited this effect through the Ang II type-1 receptor (AT1R) and ERK1/2 signaling pathway. Ang II also reduced the effect of ACE2 on ERK1/2 levels, the activity of shed ACE2, and adam17 expression in HCFs. Additionally, these Ang II-mediated reductions could be attenuated by AT1R antagonist valsartan. In conclusion, these data help to clarify how ACE2 and Ang II interact to regulate MMP-2 and control tissue remodeling in heart disease.


Asunto(s)
Angiotensina II/farmacología , Fibroblastos/efectos de los fármacos , Metaloproteinasa 2 de la Matriz/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Vasoconstrictores/farmacología , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Proteína ADAM17 , Angiotensina II/metabolismo , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Enzima Convertidora de Angiotensina 2 , Fibroblastos/citología , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Vectores Genéticos , Humanos , Lentivirus/genética , Metaloproteinasa 2 de la Matriz/genética , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Miocardio/citología , Miocardio/metabolismo , Peptidil-Dipeptidasa A/genética , Cultivo Primario de Células , Receptor de Angiotensina Tipo 1/genética , Receptor de Angiotensina Tipo 1/metabolismo , Transducción de Señal , Tetrazoles/farmacología , Valina/análogos & derivados , Valina/farmacología , Valsartán , Vasoconstrictores/metabolismo
15.
Tumour Biol ; 34(2): 837-46, 2013 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-23233043

RESUMEN

Our previous study has demonstrated that protease-activated receptor 2 (PAR2) activation mediated by tissue factor (TF)/VIIa complex triggers the ERK1/2/NF-κB signaling pathway, which further contributes to the proliferation and migration of colon cancer cell line SW620. However, the detailed mechanisms remain unclear. This study was to investigate whether protein kinase Cα (PKCα) is involved in these events and the possible mechanism. The results revealed that PAR2-activating peptide or VIIa could induce time-dependent upregulation of PKCα phosphorylation in SW620 cells and PKCα translocation from the cytoplasm to the perinuclear region and nucleus. The activation of PKCα was sufficient to induce ERK1/2 and NF-κB phosphorylation. The VIIa effect was obviously blocked by both anti-TF and anti-PAR2 antibodies. The PKCα inhibitor, safingol, inhibited ERK1/2 phosphorylation and NF-κB activation that is induced by VIIa and abrogated the enhanced proliferation, migration, and survival of SW620 cells by VIIa treatment. Both safingol and PDTC (NF-κB inhibitor) could apparently rescue the effects of VIIa on expression of MMP-9, caspase-3, TF, and Bcl-2/bax in SW620 cells. Collectively, the data in this study suggest that TF/VIIa/PAR2-induced SW620 cell proliferation, migration, and survival are ascribed to the activation of PKCα, and these effects are achieved through PKCα downstream signaling pathways, ERK1/2 and NF-κB.


Asunto(s)
Apoptosis , Movimiento Celular , Proliferación Celular , Neoplasias del Colon/patología , Factor VIIa/metabolismo , Proteína Quinasa C-alfa/metabolismo , Receptor PAR-2/metabolismo , Tromboplastina/metabolismo , Western Blotting , Adhesión Celular , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Factor VIIa/genética , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Sistema de Señalización de MAP Quinasas , FN-kappa B/genética , FN-kappa B/metabolismo , Proteína Quinasa C-alfa/antagonistas & inhibidores , Proteína Quinasa C-alfa/genética , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor PAR-2/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tromboplastina/genética , Células Tumorales Cultivadas
16.
Tumour Biol ; 34(5): 2573-81, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23616010

RESUMEN

Our previous study has demonstrated that tissue factor-factor VIIa (TF/FVIIa) complex promotes the proliferation and migration of colon cancer cell line SW620 through the activation of protease-activated receptor 2 (PAR2). In the current study, the underlying molecular mechanisms of TF/FVIIa/PAR2 signaling in SW620 cells were further explored, with the focus on the role of activator protein-1 (AP-1) subunit c-Jun. The results revealed that PAR2-AP and FVIIa could upregulate c-Jun expression and c-Jun phosphorylation in SW620 cells in a time-dependent manner. The effect of FVIIa was significantly blocked by anti-TF and anti-PAR2 antibodies. Protein kinase Cα (PKCα) inhibitor safingol and extracellular signal-regulated kinase 1 and 2 (ERK1/2) inhibitor U0126 abrogated the activation of c-Jun. In contrast, Ca(2+) chelators EGTA and thapsigargin, and p38MAPK inhibitor SB203580 had no effect. Suppression of c-Jun/AP-1 activation using a natural inhibitor curcumin decreased the expression of caspase-3, MMP-9, and TF, as well as the proliferation and migration of SW620 cells induced by PAR2-AP or FVIIa. Collectively, our findings suggest that c-Jun/AP-1 activation is required for TF/FVIIa/PAR2-induced SW620 cell proliferation and migration. PKCα and ERK1/2 are located upstream of c-Jun/AP-1 in this signaling pathway. Pharmacological inhibition of this pathway might be a novel strategy for colon cancer therapy.


Asunto(s)
Movimiento Celular , Proliferación Celular , Factor VIIa/fisiología , Sistema de Señalización de MAP Quinasas , Proteína Quinasa C-alfa/metabolismo , Receptor PAR-2/metabolismo , Tromboplastina/fisiología , Antineoplásicos/farmacología , Butadienos/farmacología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Neoplasias del Colon , Curcumina/farmacología , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Humanos , Nitrilos/farmacología , Proteína Quinasa C-alfa/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-jun/metabolismo , Esfingosina/análogos & derivados , Esfingosina/farmacología , Factor de Transcripción AP-1/antagonistas & inhibidores , Factor de Transcripción AP-1/metabolismo
17.
Cancer Invest ; 31(1): 7-16, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23170789

RESUMEN

Our previous study has demonstrated that TF/FVIIa and PAR2 are closely related to the proliferation and migration of colon cancer cell line SW620. However, the detailed molecular mechanisms in the process remain unclear. This study further investigated whether some important molecules (caspase-3, MMP-9 and CD44) are involved in the events. The results showed that PAR2-AP or FVIIa elicited time-dependent downregulation of caspase-3, and up-regulation of MMP-9 and CD44 in SW620 cells. The effects of FVIIa were TF-dependent and involving PAR2/MAPKs/NF-κB signal transduction pathways. Our study suggests that the links among PAR2/MAPKs/NF-κB may be blocked for effective treatments of colorectal cancers.


Asunto(s)
Caspasa 3/biosíntesis , Neoplasias del Colon/metabolismo , Factor VIIa/metabolismo , Receptores de Hialuranos/biosíntesis , Metaloproteinasa 9 de la Matriz/biosíntesis , FN-kappa B/metabolismo , Receptor PAR-2/genética , Receptor PAR-2/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Caspasa 3/genética , Caspasa 3/metabolismo , Procesos de Crecimiento Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Neoplasias del Colon/enzimología , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Regulación hacia Abajo , Factor VIIa/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Receptores de Hialuranos/genética , Receptores de Hialuranos/metabolismo , Sistema de Señalización de MAP Quinasas , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , FN-kappa B/genética , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos/genética
18.
Exp Mol Pathol ; 95(2): 206-12, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23895953

RESUMEN

Many miRNAs are highly expressed in spatiotemporal and precise tissue-specific patterns in development. Thus it is necessary to examine their expression pattern in mouse embryos. However, embryos from one pregnant mouse are more than enough for expression analysis such as RT-qPCR, which results in reluctant disposal of remaining embryos. Due to the limitation of short sampling time, it is vitally important to quickly preserve samples to ensure the RNA quality. Thus, it is necessary to develop appropriate methods to fix samples in advance. In this study, two fixatives [methanol/DMSO (4:1) and paraformaldehyde] were applied for embryo (12.5 dpc) fixation and two preservatives (methanol and 30% sucrose) were used for fixed embryo preservation. After storage for one month, the skin, skeletal muscle and brain tissues were dissected from the fixed and unembedded embryos. Total RNAs were extracted by TRIzol® reagent and measured by a spectrophotometer, then were subjected to amplify Actb, Hprt, Gapdh, Rnu6, Snord68 and miR-206-3p by RT-qPCR. Embryos fixed in methanol/DMSO and preserved in 100% methanol at -20°C were able to yield at least 349 bp amplifiable RNA. Although paraformaldehyde fixation and 30% sucrose preservation method only yielded amplicons less than 156 bp, it showed a remarkable ability in preserving small RNAs. Snord68 was expressed stably across skin, skeletal muscle and brain tissues like Rnu6, making its possibility as an internal control for qPCR data normalization. Using Snord68 and/or Rnu6 as internal control, we found that the miR-206-3p expression level in skin was about one quarter of its highest level in the skeletal muscle. Therefore, the techniques in this study would be useful for us to reasonably utilize and preserve precious samples.


Asunto(s)
Fijadores , MicroARNs/análisis , Estabilidad del ARN , Fijación del Tejido/métodos , Animales , Dimetilsulfóxido , Embrión de Mamíferos/química , Formaldehído , Metanol , Ratones , Ratones Endogámicos C57BL , Polímeros , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sacarosa
19.
Physiol Plant ; 148(4): 502-11, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23136980

RESUMEN

The negative charges of cell wall pectin molecules attributed by pectin methylesterase (PME, EC 3.1.1.11) contribute to Al binding capacity. We examined the expression profiles of 35 members of the PME gene family in the root apex of an Al-sensitive rice 'Zhefu802' under Al stress. While root elongation was inhibited by 40% after 3-h exposure to 25 µM Al, cell wall PME activity and the abundance of eight PME genes transcripts were increased. The same Al treatment which had almost no effect on root elongation of an Al-resistant rice ssp. japonica 'Nipponbare' did not change the expression patterns of these eight PME genes. However, when Al concentration was increased to 50 µM, by which the root elongation of 'Nipponbare' was inhibited by 40% too, the expression of these PME genes were also upregulated except two genes with no signal. These suggest a possible correlation between the upregulated genes and Al-induced inhibition of root elongation in rice. Furthermore, these eight PME genes behaved differently when subjected to CdCl2 and LaCl3 treatments, implying the specificity of different PME genes in response to different metal toxicities. The transgenic rice overexpressing one of these eight PME genes OsPME14 showed higher PME activity and Al content in root tip cell wall, and became more sensitive to Al stress, verifying the involvement of the specific PME gene in Al toxicity. Therefore, our results provided the molecular evidence to connect the expression of specific PME genes with the Al-induced inhibition of root elongation in rice.


Asunto(s)
Aluminio/toxicidad , Hidrolasas de Éster Carboxílico/metabolismo , Oryza/enzimología , Oryza/crecimiento & desarrollo , Raíces de Plantas/enzimología , Raíces de Plantas/crecimiento & desarrollo , Hidrolasas de Éster Carboxílico/genética , Pared Celular/efectos de los fármacos , Pared Celular/enzimología , Biología Computacional , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas/genética , Familia de Multigenes , Oryza/efectos de los fármacos , Oryza/genética , Fenotipo , Filogenia , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/genética
20.
Org Biomol Chem ; 11(36): 6071-89, 2013 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-23917398

RESUMEN

A classical but new kinetic equation to estimate activation energies of various hydride transfer reactions was developed according to transition state theory using the Morse-type free energy curves of hydride donors to release a hydride anion and hydride acceptors to capture a hydride anion and by which the activation energies of 187 typical hydride self-exchange reactions and more than thirty thousand hydride cross transfer reactions in acetonitrile were safely estimated in this work. Since the development of the kinetic equation is only on the basis of the related chemical bond changes of the hydride transfer reactants, the kinetic equation should be also suitable for proton transfer reactions, hydrogen atom transfer reactions and all the other chemical reactions involved with breaking and formation of chemical bonds. One of the most important contributions of this work is to have achieved the perfect unity of the kinetic equation and thermodynamic equation for hydride transfer reactions.


Asunto(s)
Hidrógeno/química , Protones , Cinética , Estructura Molecular , Compuestos Orgánicos/química , Termodinámica
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