RESUMO
To explore the relationship between autophagy and cell function, we investigated how PLAC8-mediated autophagy influences proliferation, apoptosis and epithelial-mesenchymal transition (EMT) in NPC. Colony formation analyses and CCK8 assays were used to assess the proliferative capacity of NPC cells. Transmission electron microscopy (TEM) was used to identify autophagosomes. Autophagic flux was monitored using the tandem monomeric RFP-GFP-tagged LC3 (tfLC3) assay. The rate of apoptosis in NPC cells was analysed by flow cytometry. Western blot analysis was used to evaluate the activation of autophagy and the signalling status of the AKT/mTOR pathway. Our study reveals that knocking out PLAC8 (koPLAC8) induces autophagy and apoptosis, while suppressing NPC cell proliferation and EMT. However, inhibition of autophagy with 3-methyladenine or by knocking down Beclin-1 reverses the cell proliferation, apoptosis and EMT influenced by koPLAC8. We find that koPLAC8 inhibits the phosphorylation of AKT and its downstream target, mTOR. Moreover, immunofluorescence and co-immunoprecipitation reveal complete PLAC8/AKT colocalization and PLAC8/AKT interaction, respectively. Furthermore, knockout of PLAC8 induced autophagy and inactivated AKT/mTOR signalling pathway of NPC xenografts. Overall, our findings demonstrate that koPLAC8 induces autophagy via the AKT/mTOR pathway, thereby inhibiting cell proliferation and EMT, and promoting apoptosis in NPC cells.
Assuntos
Autofagia/genética , Neoplasias Nasofaríngeas/etiologia , Neoplasias Nasofaríngeas/metabolismo , Proteínas/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Animais , Apoptose/genética , Proteína Beclina-1/genética , Proteína Beclina-1/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Modelos Animais de Doenças , Suscetibilidade a Doenças , Transição Epitelial-Mesenquimal/genética , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Modelos Biológicos , Neoplasias Nasofaríngeas/patologia , Proteínas/metabolismo , RNA Interferente Pequeno/genéticaRESUMO
The present study aimed to investigate the effects and mechanisms of PLAC8 on the epithelial-mesenchymal transition (EMT) of Nasopharyngeal carcinoma (NPC). The expression of PLAC8 in NPC and nasopharyngitis (NPG) tissues from 150 patients was determined using immunohistochemistry. The levels of PLAC8 in five NPC cell lines and nasopharyngeal permanent epithelial cell line were measured using western blotting. We then knocked out or overexpressed PLAC8 in CNE2 cells. Cell proliferation, wound healing, migration, and invasion assays were used to analyze the effects of PLAC8 on the proliferation, migration, and invasion in vivo and vitro. The results showed that the expression of PLAC8 was much higher in NPC tissues than in NPG tissues. The expression of PLAC8 was higher in all the cell lines than in the nasopharyngeal permanent epithelial cells. PLAC8 knockout resulted in significant decreases in cell proliferation, migration, and invasion; associated with lower protein levels of N-cadherin; and increased levels of E-cadherin. Overexpression of PLAC8 had the opposite effect. Furthermore, knockout of PLAC8 inactivated TGF-ß/SMAD signaling pathway and suppressed the growth of NPC xenografts. PLAC8 may promote the carcinogenesis and EMT of NPC via the TGF-ß/Smad pathway, which suggests that PLAC8 may be a potential biomarker for NPC.
Assuntos
Transição Epitelial-Mesenquimal/genética , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/patologia , Proteínas/genética , Transdução de Sinais , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Movimento Celular/genética , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica , Proteínas/metabolismo , Regulação para Cima/genéticaRESUMO
PURPOSE: To investigate the possible telomerase and alternative lengthening of telomeres (ALT) mechanisms influencing the apoptosis of laryngeal squamous cells. MATERIALS AND METHODS: The effects of the telomerase mechanism were observed by knockdown of human telomerase reverse transcriptase (hTERT). The ALT mechanism was induced by silencing related genes including TRF2, RAD51, and NBS1. Effects of telomerase and ALT mechanisms on tumor development were confirmed by xenograft tumors model. Tumor cell apoptosis was investigated by flow cytometry and Hoechst staining. Caspase-3 activity assay and Western blot were performed to investigate the possible mechanisms. RESULTS: After silencing ALT- and telomerase mechanism-related genes, Bax and Bcl-2 were increased, and nuclear factor (NF)-κB translocation and PI3K/Akt phosphorylation were inhibited. CONCLUSIONS: The inhibition of telomere-related genes inhibited the growth of laryngeal squamous cell carcinoma by promoting cell apoptosis via the PI3K/Akt pathway.
Assuntos
Apoptose/fisiologia , Carcinoma de Células Escamosas/fisiopatologia , Inativação Gênica , Neoplasias Laríngeas/fisiopatologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Telomerase/metabolismo , Telômero/fisiologia , Animais , Western Blotting , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Citometria de Fluxo , Neoplasias Laríngeas/patologia , Camundongos Nus , Inibidores de Fosfoinositídeo-3 Quinase , Plasmídeos , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , RNA Interferente Pequeno/fisiologia , Telomerase/genéticaRESUMO
Shikimic acid (1) is a renewable biomass which could be obtained sustainably through natural product isolation or metabolic engineering. Owing to its great potential in chemical conversion, the value-added utilization of this non-grain biomass has received much attention in recent years. Based on the established transformation route from shikimic acid (1) to methyl 3-dehydroshikimate (3-MDHS, 2) and to the multi-functionalized methyl 2-amino-3-cyanobenzofuran-5-carboxylate (3), we disclose a facile and transition metal-free method to access a series of N-substituted benzofuro[2,3-d]pyrimidine-4-amines in 63%-90% yields. The identification of these compounds as EGFR tyrosine kinase inhibitors has also been described. Among them, compound 5h exhibited the most potent inhibitory effect against EGFR tyrosine kinase with an IC50 of 1.7 nM and excellent antiproliferative activity against A431 and A549 cell lines with a GI50 of 5.1 and 12.3 µM, respectively.
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Biomassa , Receptores ErbB/antagonistas & inibidores , Nitrogênio/química , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/farmacologia , Células A549 , Técnicas de Química Sintética , Receptores ErbB/química , Humanos , Modelos Moleculares , Conformação Proteica , Inibidores de Proteínas Quinases/químicaRESUMO
PURPOSE: The purpose of this study was to investigate the clinical features of eosinophilic hyperplastic lymphogranuloma (EHLG) in the head and neck. MATERIALS AND METHODS: Collecting the patients who diagnose with EHLG by pathological examination. The EHLG patients with the masses involved regions, such as involved inguinal region, chest wall, abdominal wall, anterior superior iliac spine or clavicle, instead of head and neck were excluding. All of the participants will sign the informed consent form. The history data includes: clinical history, blood routine test, pathological examination, and recurrence will be collected. RESULTS: A total of 41 patients of EHLG were included. These patients predominantly presented as an enlarging and painless single or multiple masses with a history of repeated swelling. There were the complaint of itchy skin and pigmentation. The routine blood test showed that the percentage value of eosinophil increased in almost patients including 26 cases had raised absolute eosinophil count. The serum level of lgE was increased in 29 cases remarkably. With the methods of treatments, 36 patients received surgical excision, 3 patients accepted hormonotherapy, and another 2 patients for radiotherapy. The recurrence of EHLG was in 9 patients. CONCLUSIONS: EHLG is a rare disease. The clinical manifestation (itchy skin and pigmentation) and increased eosinophil play critical values to the diagnosis of EHLG. Confirmed diagnosis always depends on pathological examination. Surgery is a preferred treatment, while low dose of radiotherapy is necessary for preventing relapse after operation and hormonotherapy.
Assuntos
Hiperplasia Angiolinfoide com Eosinofilia/diagnóstico , Hiperplasia Angiolinfoide com Eosinofilia/terapia , Adolescente , Adulto , Idoso , Hiperplasia Angiolinfoide com Eosinofilia/complicações , Criança , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Adulto JovemRESUMO
A total synthesis of coumestrol (1) and aureol (2) is described. The Perkin condensation of 2-bromo-4-hydroxylphenylacetic acid (6) and o-hydroxybenzaldehydes (7) gave the corresponding 2'-bromo-3-arylcoumarins (9). A copper-catalyzed consecutive hydroxylation and aerobic oxidative coupling of 9 under microwave conditions facilitated the total synthesis of 1 and 2, respectively, with spectroscopic data highly similar to those of natural products.
Assuntos
Cumestrol/síntese química , Sesquiterpenos/síntese química , Catálise , Cobre/química , Cumarínicos/química , Cumestrol/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Sesquiterpenos/químicaRESUMO
A series of ebselen derivatives were designed, synthesised and evaluated as inhibitors of cholinesterases (ChEs) and glutathione peroxidase (GPx) mimics. Most of the compounds were found to be potent against AChEs and BuChE, compounds 5e and 5i, proved to be the most potent against AChE with IC50 values of 0.76 and 0.46 µM, respectively. Among these hybrids, most of the compounds were found to be good GPx mimics compare with ebselen. The selected compounds 5e and 5i were also used to determine the catalytic parameters and in vitro hydrogen peroxide scavenging activity. The results indicate that compounds 5e and 5i may be excellent multifunctional agents for the treatment of AD.
Assuntos
Acetilcolinesterase/química , Antioxidantes/síntese química , Azóis/química , Azóis/síntese química , Materiais Biocompatíveis/síntese química , Inibidores da Colinesterase/síntese química , Compostos Organosselênicos/química , Compostos Organosselênicos/síntese química , Acetilcolinesterase/metabolismo , Doença de Alzheimer/tratamento farmacológico , Antioxidantes/metabolismo , Antioxidantes/uso terapêutico , Azóis/metabolismo , Azóis/uso terapêutico , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/uso terapêutico , Butirilcolinesterase/química , Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/metabolismo , Inibidores da Colinesterase/uso terapêutico , Glutationa Peroxidase/química , Glutationa Peroxidase/metabolismo , Humanos , Isoindóis , Compostos Organosselênicos/metabolismo , Compostos Organosselênicos/uso terapêutico , Ligação ProteicaRESUMO
Our previous study found that in nasopharyngeal carcinoma (NPC) cells, overexpression of Notch2 can inhibit epithelial-mesenchymal transition (EMT), which plays a vital role in mediating radiosensitivity. The purpose of this study was to explore the radiosensitizing efficacy of the Notch2 gene in NPC cells and its potential mechanism. We used the recombinant plasmid transfection technique to establish Notch2-overexpressing 5-8 F and CNE-2 NPC cells. Cell proliferation, radiosensitivity, apoptosis and cell cycle distribution were assessed by cell counting kit-8 (CCK-8) experiments, colony formation experiments and flow cytometry. The levels of proteins related to cell cycle, apoptosis, and the AKT/mTOR signaling pathway were evaluated by using Western blotting. The results suggested that Notch2 overexpression increased the radiosensitivity of NPC cells, with sensitizing enhancement ratios (SERs) of 1.24 (5-8 F cells) and 1.34 (CNE-2 cells). Flow cytometry indicated that the level of apoptosis and percentage of cells in G2/M-phase were highest in NPC cells overexpressing Notch2 and treated with radiotherapy compared to cells overexpressing Notch2 alone or administered radiotherapy alone. Western blotting showed that compared to that of cells treated with Notch2 overexpression or radiotherapy alone, the levels of γH2AX, Bax, Bcl-2, Cyclin D1 and AKT/mTOR signaling pathway-related proteins were modified in NPC cells overexpressing Notch2 and treated with radiotherapy. These findings showed that overexpression of Notch2 can increase the radiosensitivity of NPC cells by inhibiting the AKT/mTOR pathway.AbbreviationsNPC: Nasopharyngeal carcinoma; EMT: Epithelial-mesenchymal transition; CCK8: Cell counting kit-8; EBV: Epstein-Barr virus; FBS: Fetal bovine serum; PE: Plating efficiency; SF: Survival fraction; SER: Sensitizing enhancement ratio; DSBs: DNA double-strand breaks[Figure: see text].
Assuntos
Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas , Proteínas Proto-Oncogênicas c-akt/genética , Tolerância a Radiação/genética , Receptor Notch2/genética , Linhagem Celular Tumoral , Humanos , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/metabolismo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor Notch2/metabolismo , Transdução de Sinais/genética , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
BACKGROUND: Centromere protein N (CENP-N) has been reported to be highly expressed in malignancies, but its role and mechanism in nasopharyngeal carcinoma (NPC) are unknown. METHODS: Abnormal CENP-N expression from NPC microarrays of GEO database was analyzed. CENP-N expression level was confirmed in NPC tissues and cell lines. Stable CENP-N knockdown and overexpression NPC cell lines were established, and transcriptome sequencing after CENP-N knockdown was performed. In vitro and in vivo experiments were performed to test the impact of CENP-N knockdown in NPC cells. ChIP and dual luciferase reporter assays were used to verify the combination of IRF2 and CENP-N. Western blot analysis, cellular immunofluorescence, immunoprecipitation and GST pulldown assays were used to verify the combination of CENP-N and AKT. RESULTS: CENP-N was confirmed to be aberrantly highly expressed in NPC tissues and cell lines and to be associated with high 18F-FDG uptake in cancer nests and poor patient prognosis. Transcriptome sequencing after CENP-N knockdown revealed that genes with altered expression were enriched in pathways related to glucose metabolism, cell cycle regulation. CENP-N knockdown inhibited glucose metabolism, cell proliferation, cell cycling and promoted apoptosis. IRF2 is a transcription factor for CENP-N and directly promotes CENP-N expression in NPC cells. CENP-N affects the glucose metabolism, proliferation, cell cycling and apoptosis of NPC cells in vitro and in vivo through the AKT pathway. CENP-N formed a complex with AKT in NPC cells. Both an AKT inhibitor (MK-2206) and a LDHA inhibitor (GSK2837808A) blocked the effect of CENP-N overexpression on NPC cells by promoting aerobic glycolysis, proliferation, cell cycling and apoptosis resistance. CONCLUSIONS: The IRF2/CENP-N/AKT axis promotes malignant biological behaviors in NPC cells by increasing aerobic glycolysis, and the IRF2/CENP-N/AKT signaling axis is expected to be a new target for NPC therapy.
Assuntos
Proteínas Cromossômicas não Histona/metabolismo , Fator Regulador 2 de Interferon/metabolismo , Neoplasias Nasofaríngeas/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Apoptose , Ciclo Celular , Proliferação de Células , Genes Sintéticos , Humanos , Camundongos , Camundongos Nus , Prognóstico , Proteínas Recombinantes , Transdução de Sinais , Efeito Warburg em OncologiaRESUMO
ABSTRACT: MicroRNAs (miRNAs) play critical roles in carcinogenesis and development of cancers. In this study, we analyzed the eccentrically expressed miRNAs in head and neck squamous cell carcinoma (HNSCC) tissues based on the miRNA-Seq data of HNSCC patients available in the Cancer Genome Atlas database. Aberrant expression of 2589 miRNAs was detected in HNSCC tissues (1128 downregulated and 1461 upregulated). The differential expression levels of the miRNAs were further validated by analysis of 25 HNSCC samples and paired control tissues and compared with the Gene Expression Omnibus database to determine the candidate miRNAs. Quantitative reverse transcription polymerase chain reaction was used to compare the expression of these candidate miRNAs between 22 fresh HNSCC tissue samples and 11 control samples. In addition, the relationship between the expression of these candidate miRNAs and Tumor, Node, Metastases staging of HNSCC was analyzed. Compared with the expression in control tissues, the levels of hsa-miR-410-3p, hsa-miR-411-5p, hsa-miR-125b-2-3p, and hsa-miR-99a-3p were significantly lower in HNSCC. According to the Cancer Genome Atlas dataset analyzed, all 4 miRNAs were shown to inhibit tumor progression (T stage), positive lymph node metastasis (N stage), and distant metastasis (M stage) in HNSCC. Kyoto Encyclopedia of Genes and Genomes analysis showed that genes regulated by these 4 miRNAs were enriched in certain pathways, including the transforming growth factor-ß signaling pathway and the Hippo pathway. Enriched gene ontology terms mainly included regulation of transcription, cell proliferation, and apoptosis, which are well-characterized functions of miRNAs. Moreover, all 4 miRNAs inhibited the progression of primary tumors (T stage) and metastasis of regional lymph nodes (N stage). The top 4 aberrantly expressed miRNAs identified in this study have great clinical value in developing strategies for early diagnosis and treatment of HNSCC. More intensive studies are required to elucidate the mechanism underlying the roles of these miRNAs in HNSCC.
Assuntos
Neoplasias de Cabeça e Pescoço/genética , MicroRNAs/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Idoso , Biomarcadores Tumorais/genética , Regulação para Baixo , Feminino , Regulação Neoplásica da Expressão Gênica , Neoplasias de Cabeça e Pescoço/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma de Células Escamosas de Cabeça e Pescoço/sangueRESUMO
In vitro cell experiments showed that knocking out the placenta-specific protein 8 (PLAC8) gene significantly increased the sensitivity of tumor cells to radiation. This study used two nude mouse models of nasopharyngeal carcinoma (NPC) to investigate the radio-sensitization and molecular mechanism of PLAC8 knockout in vivo. The expression of PLAC8 in 120 NPC tissues and 30 nasopharyngitis (NPG) tissues was detected by immunohistochemistry (IHC) to analyze the relationship between PLAC8 and neck lymph node metastasis and prognosis in NPC patients. The mRNA expression level of PLAC8 in several NPC cell lines was detected by semi-quantitative RT-PCR. The PLAC8 gene was knocked out in CNE-2 cells using CRISPR/Cas9. The effect of PLAC8 gene knockout on the radiotherapy sensitivity of NPC cells was analyzed by establishing model 1 and model 2 tumor-bearing nude mouse models with two different irradiation methods. The expression of γH2AX, Bax, Bcl-2, Caspase-3 and cleaved Caspase-3 was detected by immunofluorescence (IF), IHC and western blot analysis. PLAC8 expression was significantly increased in NPC tissue samples and NPC cell lines compared with NPG tissue samples and normal cell lines (P<0.01). PLAC8 upregulation was associated with lymph node metastasis and a poor prognosis in patients with NPC (P<0.01). Both animal models showed that radiotherapy after PLAC8 knockout significantly slowed tumor growth and reduced tumor volume, with tumor inhibition rates of 100% and 66.04%, respectively. In model 2, PLAC8 knockout with radiotherapy increased the expressions of γH2AX, Bax, Caspase-3 and cleaved Caspase-3 but decreased the expression of Bcl-2 (P<0.01). In model 1, there was no tumor formation at the site where the cancer cells were injected. The expression levels of γH2AX, Bax, Caspase-3 and cleaved Caspase-3 in skin tissues taken at the injection site were lower than those in NPC tissues treated with radiotherapy, while the expression level of Bcl-2 was higher (P<0.01). PLAC8 expression is closely related to neck metastasis and the prognosis of NPC. PLAC8 gene knockout significantly increases the radio-sensitivity of NPC cells in vivo by promoting apoptosis, which is an effective strategy for the radiotherapy sensitization of NPC.
RESUMO
A copper-catalyzed intramolecular cross dehydrogenative C-O coupling reaction of 2'-hydroxyl-3-arylcoumarins was developed. This protocol provided a facile and efficient strategy for the construction of natural coumestans and derivatives in moderate to high yields. This transformation exhibited good functional group compatibility and was amenable to substrates with free phenolic hydroxyl groups.
RESUMO
BACKGROUND: Clinically, distant metastasis after primary treatment remains a key problem in nasopharyngeal carcinoma (NPC). Thus, identification of the underlying mechanisms and development of novel therapeutic strategies are urgently needed. NOTCH has been shown to function as a tumor promotor that enhances angiogenesis, cancer invasion and metastasis in NPC. However, the precise roles of the four individual NOTCH receptors and their mechanisms of action are unclear. METHODS: We used Western blot analysis, immunofluorescence, immunohistochemical analysis, phalloidin staining, mouse tumor metastatic dissemination models, gene set enrichment analysis, immunoprecipitation assays and a series of functional assays to determine the potential role of NOTCH2 in regulating NPC metastasis. RESULTS: NOTCH2 expression in the NPC tissues of patients with cervical lymph node metastasis was lower than that of patients without cervical lymph node metastasis. Correspondingly, NOTCH2 expression was low in metastatic and poorly differentiated NPC cells. NOTCH2 expression correlated negatively with survival time in patients with NPC. Suppression of NOTCH2 expression promoted NPC cell metastasis, whereas NOTCH2 overexpression inhibited this process. Furthermore, NOTCH2 attenuated the TRAF6-AKT signaling axis via an interaction between the NOTCH2 intracellular domain (N2ICD) and TRAF6, which inhibited epithelial-mesenchymal transition (EMT) and eventually suppressed NPC metastasis. CONCLUSIONS: These findings reveal that loss of NOTCH2 activates the TRAF6/AKT axis and promotes metastasis in NPC, suggesting that NOTCH2 may represent a therapeutic target for the treatment of NPC.
Assuntos
Transição Epitelial-Mesenquimal , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor Notch2/metabolismo , Fator 6 Associado a Receptor de TNF/metabolismo , Adulto , Idoso , Animais , Biomarcadores , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Pessoa de Meia-Idade , Modelos Moleculares , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/mortalidade , Metástase Neoplásica , Estadiamento de Neoplasias , Prognóstico , Ligação Proteica , Receptor Notch2/genética , Transdução de SinaisRESUMO
Many plant-specialized metabolites have remedial properties and provide an endless chemical resource for drug discovery. However, most of these metabolites have promiscuous binding targets in mammalian cells and elicit a series of responses that collectively change the physiology of the cells. To explore the potential of these multi-functional and multi-targeted drugs, it is critical to understand the direct relationships between their key chemical features, the corresponding binding targets and the relevant biological effects, which is a prerequisite for future drug modification and optimization. Methods: We introduced and demonstrated a general workflow, called Comparative Profiling of Analog Targets (CPAT), to connect specific biological effects with defined chemical structures of drugs. Using resveratrol (RSV) as an example, we have synthesized and characterized a series of partial functional analogs of RSV. An analog (named RSVN) that specifically lost the inhibitory effect of RSV in cell migration was identified. The binding targets of RSVN and RSV was profiled and compared. Results: Comparative profiling of the RSV and RSVN binding targets showed that, unlike RSV, RSVN failed to target specific components involved in DNA methylation (histone deacetylase 1 [HDAC1] and DNA methyltransferase 3 alpha [DNMT3a]), suggesting that RSV suppresses cell migration through epigenetic regulation. Indeed, RSV treatment recruited HDAC1 and DNMT3a to the promoter region of the focal adhesion kinase (FAK), a key factor involved in cell adhesion, enhanced the promoter methylation, and thus attenuated the protein expression. The inhibitory effect of RSV in cell migration was diminished once FAK expression was restored. Thus, the mechanism of RSV in inhibiting cell migration could be largely accounted to epigenetically control of FAK expression. Conclusion: Our results showed that even though RSV exhibits promiscuous binding, its inhibitory effect on cell migration can be mechanistically understood. First, the presence of 4'-hydroxystilbene within the RSV structure is essential for this activity. Second, it inhibits cell migration through epigenetically based downregulation of FAK expression. Taken together, we propose that CPAT might also be adapted to delineate the specific function of other natural products (NPs) that exhibit binding promiscuity.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Perfilação da Expressão Gênica , Melanoma/tratamento farmacológico , Metástase Neoplásica/tratamento farmacológico , Resveratrol/farmacologia , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Movimento Celular/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Resveratrol/administração & dosagem , Resveratrol/análogos & derivados , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: To study the anatomic characteristics of recurrent laryngeal nerve during thyroid surgery. METHOD: A retrospective review of surgical data of 307 patients undertook thyroid surgery was conducted. RESULT: Total 342 recurrent laryngeal nerves were identified during the surgery(184 on the right side, left 158). 215 (62.9%) nerves were deep to the inferior thyroid artery, 106(31.0%)were superficial to the artery, 21(7.5%) were between the arterial branches. A nerve bifurcation was found in 203(59.4%). None of nerve bifurcation was found in 136(39.8%). 3(0.9%)were confirmed to hold non-recurrent laryngeal nerves during operations. No patient showed permanent laryngeal recurrent nerve paralysis postoperatively. CONCLUSION: The careful dissection and protection of the recurrent laryngeal nerve was an effective method to prevent its injury during thyroid surgery.
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Doenças dos Nervos Cranianos/etiologia , Nervo Laríngeo Recorrente , Glândula Tireoide/cirurgia , Artérias , Doenças dos Nervos Cranianos/prevenção & controle , Dissecação , Humanos , Período Pós-Operatório , Estudos Retrospectivos , Tireoidectomia , Paralisia das Pregas VocaisRESUMO
A novel series of compounds obtained by fusing the cholinesterase inhibitor donepezil and the antioxidant ebselen were designed as multi-target-directed ligands against Alzheimer's disease. An in vitro assay showed that some of these molecules did not exhibit highly potent cholinesterase inhibitory activity but did have various other ebselen-related pharmacological effects. Among the molecules, compound 7d, one of the most potent acetylcholinesterase inhibitors (IC50 values of 0.042 µM for Electrophorus electricus acetylcholinesterase and 0.097 µM for human acetylcholinesterase), was found to be a strong butyrylcholinesterase inhibitor (IC50 = 1.586 µM), to possess rapid H2O2 and peroxynitrite scavenging activity and glutathione peroxidase-like activity (ν0 = 123.5 µM min(-1)), and to be a substrate of mammalian TrxR. A toxicity test in mice showed no acute toxicity at doses of up to 2000 mg/kg. According to an in vitro blood-brain barrier model, 7d is able to penetrate the central nervous system.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Azóis/síntese química , Azóis/farmacologia , Indanos/síntese química , Indanos/farmacologia , Terapia de Alvo Molecular , Compostos Organosselênicos/síntese química , Compostos Organosselênicos/farmacologia , Piperidinas/síntese química , Piperidinas/farmacologia , Acetilcolinesterase/química , Acetilcolinesterase/metabolismo , Peptídeos beta-Amiloides/química , Animais , Azóis/metabolismo , Azóis/toxicidade , Butirilcolinesterase/metabolismo , Técnicas de Química Sintética , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/metabolismo , Inibidores da Colinesterase/farmacologia , Inibidores da Colinesterase/toxicidade , Donepezila , Electrophorus , Glutationa Peroxidase/metabolismo , Humanos , Indanos/metabolismo , Indanos/toxicidade , Isoindóis , Cinética , Ligantes , Camundongos , Simulação de Acoplamento Molecular , Compostos Organosselênicos/metabolismo , Compostos Organosselênicos/toxicidade , Fragmentos de Peptídeos/química , Ácido Peroxinitroso/metabolismo , Piperidinas/metabolismo , Piperidinas/toxicidade , Multimerização Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína , Tiorredoxina Dissulfeto Redutase/metabolismo , Testes de Toxicidade AgudaRESUMO
We investigated the relationship between telomerase activity and the expression of human telomerase reverse transcriptase (hTERT) and prostate apoptosis response 4 (Par-4) proteins in laryngocarcinoma cells. Following treatment with short hairpin RNA (shRNA) against hTERT, hTERT protein expression was inhibited, while Par-4 expression was increased. These changes were statistically significant (p<0.05), with Par-4 and hTERT expression being negatively correlated (p<0.05, r=-0.908). However, we found no correlation between telomerase activity and hTERT expression. Our findings suggest that hTERT exerts its anti-apoptotic effects through a telomerase-independent pathway, which is likely to intersect with the Par-4 apoptosis pathway.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Neoplasias Laríngeas/patologia , Interferência de RNA , Telomerase/genética , Apoptose/genética , Linhagem Celular Tumoral , Humanos , Neoplasias Laríngeas/metabolismo , Masculino , Telomerase/antagonistas & inibidores , Telomerase/metabolismoRESUMO
OBJECTIVE: To analyze the characteristic of chronic paranasal sinusitis CT scans and discuss the relationship between the anatomic variations in ostiomeatal complex (OMC) and the incidence of chronic sinusitis. METHOD: One hundred and ninety-seven cases diagnosis as sinusitis were selected and analyzed,whose age ranged between 4 and 74 years old. Several common anatomical abnormalities in ostiomeatal complex were evaluated through the observation of the coronal and axial plane CT scans of the paranasal sinusitis. RESULT: The incidence of anatomic variations in OMC concentrated in the age between 11-20. The incidence of chronic sinusitis also concentrated in the age between 11-20. Deviation of nasal septum, uncinate variations, inferior-turbinate hypertrophy were observed between the chronic sinusitis group and the none chronic sinusitis group,the correlation between deviation of nasal septum, uncinate variations, inferior-turbinate hypertrophy and chronic sinusitis were significant (P<0.05). CONCLUSION: The occurrence of OMC variation is common. The anatomic variations may be one of the causes of chronic paranasal sinusitis, especially deviation of nasal septum, uncinate variations and inferior-turbinate hypertrophy. If these abnormalities were altered,it will have a great significance for the preservation and treatment of chronic paranasal sinusitis.
Assuntos
Sinusite/diagnóstico por imagem , Tomografia Computadorizada Espiral , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Septo Nasal/diagnóstico por imagem , Seios Paranasais/diagnóstico por imagem , Adulto JovemRESUMO
OBJECTIVE: To explore the effect of a comprehensive surgical approach of hyoid suspension with Repose system plus uvulopalatopharyngoplasty (UPPP) on the treatment of severe obstructive sleep apnea hypopnea syndrome (OSAHS). METHOD: Twelve patients with severe OSAHS (AHI > 40) diagnosed by polysomnography, were determined for the obstruction in both the velo-pharyngeal and the hypopharynx by preoperative physical examination, fiberoptic pharyngo-laryngoscopy. Two cases were given continuous airway pressure via nose, seven days before operation and then were performed hyoid suspension plus UPPP. The follow up was 3 months postoperatively. The pair T test was used to compare the preoperative and postoperative results by SPSS11.0 for windows. RESULT: At postoperative 3 month, there were statistically significance between preoperative and postoperative measurement in all. Mean AHI decreased from preoperative 69.28 +/- 6.50 to postoperative 19.77 +/- 9.23, lowest mean oxygen saturation increased from 65.25 -/+ 3.14 to 90.17 +/- 2.86. Snoring and sleep apnea disappeared or lessened, with the disappearance or decrease of lethargy at daytime. No severe complication occurred. CONCLUSION: Comprehensive surgical approach of hyoid suspension with Repose system plus UPPP is effective surgical approach for patients with severe OSAHS who suffered from oropharyngeal and hypopharyngeal obstruction in short- term result.
Assuntos
Osso Hioide/cirurgia , Procedimentos Cirúrgicos Otorrinolaringológicos/métodos , Apneia Obstrutiva do Sono/cirurgia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Palato Mole/cirurgia , Faringe/cirurgia , Resultado do Tratamento , Úvula/cirurgia , Adulto JovemRESUMO
OBJECTIVE: To study the effect of radiofrequency ablation on perennial allergic rhinitis. METHOD: Thirty-five cases of perennial allergic rhinitis undergone the treatment in radiofrequency ablation were study. RESULT: All patients were followed more than 6 months, total effective rate is 94.3% (33/35). Among the total, excellence is 85.7% (30/35). CONCLUSION: The radiofrequency ablation is a simple and safe instrument to treat perennial allergic rhinitis.