RESUMO
The sex-determining region Y-box 4 (SOX4), a transcription factor, is involved in various developmental processes. It has been reported in multiple human cancers. However, the prognostic value and its exact role in chondrosarcoma remain poorly understood. In the current study, SOX4 was overexpressed in 28 of 92 (30.4 %) interpretable chondrosarcoma patients compared with 3 of 43 (6.9 %) interpretable chondroma cases (P = 0.003). Its overexpression in chondrosarcoma was significantly associated with histological grade (P < 0.001) and the presence of tumor recurrence (P = 0.041). In addition, SOX4 overexpression was notably correlated with c-MYC (P = 0.011) and P53 (P = 0.029) expression as well as high Ki67 labeling index (LI) (P < 0.001) in our cohort. More importantly, we found that SOX4 was an unfavorable independent prognostic factor for chondrosarcoma patients with low histological grade. Functionally, SOX4 silencing significantly suppressed the proliferation, migratory, and invasive capacity of SW1353 cells, suggesting an oncogenic role of SOX4 in chondrosarcoma in vitro. In an attempt of characterizing SOX4 overexpression mechanism, we identified miR-30a as a tumor suppressor that directly targets SOX4 in chondrosarcoma cells. Clinically, miR-30a expression was negatively correlated with SOX4 expression in chondrosarcoma cases. In all, we identified that SOX4 was oncogenic in chondrosarcoma and negatively regulated by miR-30a in vitro. Importantly, SOX4 overexpression may serve as a prognostic marker for patients with low-histological-grade chondrosarcoma.
Assuntos
Neoplasias Ósseas/genética , Condrossarcoma/genética , MicroRNAs/genética , Recidiva Local de Neoplasia/genética , Fatores de Transcrição SOXC/biossíntese , Adulto , Idoso , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Condrossarcoma/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Recidiva Local de Neoplasia/patologia , Prognóstico , Fatores de Transcrição SOXC/genéticaRESUMO
Psoriasis is a skin disease with autoimmune tendency, and taxifolin is an effective flavonoid with anti-inflammatory activity. It has been reported that taxifolin alleviates psoriatic dermatitis, but the detailed regulatory mechanism of keratinocyte proliferation is unclear. In this study, we revealed the mechanism of taxifolin on imiquimod-induced inflammatory infiltration and keratinocyte over-proliferation. Our results show that taxifolin prevented proliferation cycle of keratinocyte in a concentration-dependent manner. Over-proliferation and abnormal apoptosis of epidermal cells were obvious in the mouse model of psoriasis induced by imiquimod. Taxifolin treatment improved erythema and scales of psoriatic lesions in mice, and reduced the proportion of CD3 + cells, especially γδT cells, in lesions and thymus. Therefore, taxifolin decreased the expression level of IL-17A-dominated inflammatory cytokines. Proteomic analysis showed that 30 up-regulated proteins and 23 down-regulated proteins were compared with the lesions before and after the treatment with taxifolin. Among them, cytoplasmic phospholipase A2 (cPLA2), the key enzyme of the pro-inflammatory mediator, was the most significantly down-regulated protein. And enriched KEGG pathway shown that PPAR-γ pathway was most involved. Taxifolin significantly reduced p-cPLA2 and increased PPAR-γ protein level in keratinocytes and lesions induced by IL-17 and imiquimod respectively. Meanwhile, phosphorylation of ERK and P-38 were also inhibited. These results suggest that taxifolin prevented imiquimode-induced excessive immune activation and keratinocyte proliferation by decreasing p-cPLA2 and regulating the PPAR-γ pathway. Our study provides new insights into the cellular regulatory mechanisms of taxifolin in psoriasis.
Assuntos
Psoríase/tratamento farmacológico , Quercetina/análogos & derivados , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citoplasma/metabolismo , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Humanos , Imiquimode , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Masculino , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/metabolismo , PPAR gama/metabolismo , Fosfolipases A2/metabolismo , Fosforilação/efeitos dos fármacos , Proteômica , Psoríase/induzido quimicamente , Psoríase/metabolismo , Quercetina/farmacologia , Quercetina/uso terapêutico , Transdução de Sinais/efeitos dos fármacosRESUMO
Dauricine, a bioactive component of Asiatic Moonseed Rhizome, has been widely used to treat a large number of inflammatory diseases in traditional Chinese medicine. In our study, we demonstrated that dauricine inhibited colon cancer cell proliferation and invasion, and induced apoptosis by suppressing nuclear factor-kappaB (NF-kappaB) activation in a dose- and time-dependent manner. Addition of dauricine inhibited the phosphorylation and degradation of IkappaBalpha, and the phosphorylation and translocation of p65. Moreover, dauricine down-regulated the expression of various NF-kappaB-regulated genes, including genes involved cell proliferation (cyclinD1, COX2, and c-Myc), anti-apoptosis (survivin, Bcl-2, XIAP, and IAP1), invasion (MMP-9 and ICAM-1), and angiogenesis (VEGF). In athymic nu/nu mouse model, we further demonstrated that dauricine significantly suppressed colonic tumor growth. Taken together, our results demonstrated that dauricine inhibited colon cancer cell proliferation, invasion, and induced cell apoptosis by suppressing NF-kappaB activity and the expression profile of its downstream genes. These findings provide evidence for a novel role of dauricine in preventing or treating colon cancer through modulation of NF-kappaB singling pathway.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Apoptose/efeitos dos fármacos , Benzilisoquinolinas/farmacologia , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Tetra-Hidroisoquinolinas/farmacologia , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/uso terapêutico , Benzilisoquinolinas/química , Benzilisoquinolinas/uso terapêutico , Caspases/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Neoplasias do Colo/tratamento farmacológico , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas I-kappa B/metabolismo , Camundongos , Proteína Quinase 8 Ativada por Mitógeno/metabolismo , Proteína Quinase 9 Ativada por Mitógeno/metabolismo , Estrutura Molecular , Inibidor de NF-kappaB alfa , Invasividade Neoplásica , Transplante de Neoplasias , Poli(ADP-Ribose) Polimerases/metabolismo , Tetra-Hidroisoquinolinas/química , Tetra-Hidroisoquinolinas/uso terapêutico , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
BACKGROUND: Tumor necrosis factor alpha (TNFalpha) has been used to treat certain tumors in clinic trials. However, the curative effect of TNFalpha has been undermined by the induced-NF-kappaB activation in many types of tumor. Maslinic acid (MA), a pharmacological safe natural product, has been known for its important effects as anti-oxidant, anti-inflammatory, and anti-viral activities. The aim of this study was to determine whether MA potentiates the anti-tumor activity of TNFalpha though the regulation of NF-kappaB activation. RESULTS: In this study, we demonstrate that MA significantly enhanced TNFalpha-induced inhibition of pancreatic cancer cell proliferation, invasion, and potentiated TNFalpha-induced cell apoptosis by suppressing TNFalpha-induced NF-kappaB activation in a dose- and time-dependent manner. Addition of MA inhibited TNFalpha-induced IkappaBalpha degradation, p65 phosphorylation, and nuclear translocation. Furthermore, MA decreased the expression levels of NF-kappaB-regulated genes, including genes involved in tumor cell proliferation (Cyclin D1, COX-2 and c-Myc), apoptosis (Survivin, Bcl-2, Bcl-xl, XIAP, IAP-1), invasion (MMP-9 and ICAM-1), and angiogenesis (VEGF). In athymic nu/nu mouse model, we further demonstrated that MA significantly suppressed pancreatic tumor growth, induced tumor apoptosis, and inhibited NF-kappaB-regulated anti-apoptotic gene expression, such as Survivin and Bcl-xl. CONCLUSIONS: Our data demonstrate that MA can potentiate the anti-tumor activities of TNFalpha and inhibit pancreatic tumor growth and invasion by activating caspase-dependent apoptotic pathway and by suppressing NF-kappaB activation and its downstream gene expression. Therefore, MA together with TNFalpha could be new promising agents in the treatment of pancreatic cancer.
Assuntos
Antineoplásicos/administração & dosagem , NF-kappa B/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Triterpenos/administração & dosagem , Fator de Necrose Tumoral alfa/administração & dosagem , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Ensaio de Desvio de Mobilidade Eletroforética , Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Masculino , Camundongos , Camundongos Nus , NF-kappa B/biossíntese , Análise de Sequência com Séries de Oligonucleotídeos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
AIMS: To evaluate the therapeutic benefits of Hesperidin (Hes) using an imiquimod (IMQ)-induced psoriasis-like mouse model and human immortalized keratinocytes (HaCaT) cells stimulated with lipopolysaccharide (LPS). METHODS: Mice were treated with IMQ and orally administered Hes (125-500â¯mg/kg/day), methotrexate (MTX) 1â¯mg/kg/day or distilled water. HaCaT cells were stimulated with LPS (1⯵g/mL) and relevant indices were measured after administration with different concentrations of Hes (5-20⯵g/mL) for 24â¯h. Inflammatory skin lesions in IMQ mice were evaluated using the psoriasis area severity index (PASI) and pathological staining. Proteins in the IRS-1/ERK1/2 pathway and inflammatory factors were assessed using western blotting or quantitative real-time PCR. In addition, factors related to IRS-1 secretion levels were assessed by enzyme-linked immunosorbent assays. Extracellular flux (XF) analysis was used to assess cellular metabolic levels. KEY FINDINGS: Hes significantly improved psoriasis-like skin lesions of IMQ-treated mice and inhibited LPS-induced HaCaT cell proliferation. In addition, Hes remarkably decreased PASI scores, reduced epidermal thickness, decreased proliferation and differentiation of epidermal cells, inhibited mRNA expression of inflammatory factors, reduced local skin lesions and serum insulin and glucose levels. Furthermore, Hes modulated the secretion levels of serum Leptin, Adiponectin and Resistin, and inhibited the activation of the IRS-1/ERK1/2 signaling pathway and regulated HaCaT cells metabolism. SIGNIFICANCE: This study demonstrated that Hes administration could have significant therapeutic value for the prevention and clinical treatment of psoriasis.
Assuntos
Fármacos Dermatológicos/uso terapêutico , Hesperidina/uso terapêutico , Imiquimode/farmacologia , Queratinócitos/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Psoríase/tratamento farmacológico , Animais , Western Blotting , Proliferação de Células/efeitos dos fármacos , Fármacos Dermatológicos/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Hesperidina/farmacologia , Humanos , Queratinócitos/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Metotrexato/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Psoríase/induzido quimicamente , Psoríase/metabolismo , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Our previous studies suggested that paeonol, the active constituent of the traditional Chinese medicine Cortex Moutan, may be an effective treatment for inflammatory disorders. In the present study, the therapeutic potential of paeonol on atopic dermatitis (AD) was investigated using animal and cell experiments. ADlike lesions were induced by repeated application of 1chloro2,4dinitrobenzene (DNCB) to the shaved dorsal skin of BALB/c mice, and P815 cells were used for in vitro assays. The skin lesions, serum and spleens of the mice were analyzed using lesion severity scoring, histological analysis, flow cytometry, reverse transcriptionquantitative polymerase chain reaction, western blotting and ELISA, in order to investigate the antiAD effects of paeonol. In addition, western blotting and ELISA were conducted for in vitro analysis of P815 cells. The results demonstrated that oral administration of paeonol inhibited the development of DNCBinduced ADlike lesions in the BALB/c mice by reducing severity of the lesions, epidermal thickness and mast cell infiltration; this was accompanied by reduced levels of immunoglobulin E and inflammatory cytokines [interleukin (IL)4, histamine, IL13, IL31 and thymic stromal lymphopoietin], along with regulation of the T helper (Th) cell subset (Th1/Th2) ratio. Application of paeonol also reduced the protein expression levels of phosphorylated (p)p38 and pextracellular signalregulated kinase (ERK) in skin lesions. In vitro, paeonol reduced the expression levels of tumor necrosis factorα and histamine in P815 cells, and inhibited p38/ERK/mitogenactivated protein kinase signaling. The present findings indicated that paeonol may relieve dermatitis by acting on cluster of differentiation 4+ T and mast cells; therefore, paeonol may represent a potential therapeutic strategy for the treatment of allergic inflammatory conditions via immunoregulation.
Assuntos
Acetofenonas/farmacologia , Dermatite Atópica/etiologia , Dermatite Atópica/metabolismo , Dinitroclorobenzeno/efeitos adversos , Mastócitos/imunologia , Mastócitos/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Animais , Biomarcadores , Linhagem Celular , Citocinas/genética , Citocinas/metabolismo , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/patologia , Modelos Animais de Doenças , Feminino , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Mediadores da Inflamação/metabolismo , Camundongos , Transdução de Sinais , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismoRESUMO
Matrine, is a bioactive compound isolated from Sophora flavescens (Ku shen), an herb used in Chinese traditional medicine that possesses wide-reaching pharmacological action. Psoriasis is a chronic relapsing inflammatory disorder with an incompletely understood pathophysiology, and dendritic cells (DCs) play a central role in the disease. This study aimed to explore DCs related potential mechanisms based on the effect of matrine on imiquimod (IMQ)-induced psoriasiform dermatitis in BALB/c mice and DCs simulated by resiquimod. Mice with IMQ-induced psoriasiform cutaneous lesions were treated with matrine [12.5, 25 or 50 mg/(kg·d), for 6 days]. Morphology, histological changes, keratinocyte proliferation and differentiation, inflammatory cell infiltration, protein expression levels of myeloid differentiation factor 88 (MyD88), and mRNA expression levels of inflammatory factors [interleukin (IL)-12, IL-23, and IL-1ß] in lesional skin were assessed. The application of matrine decreased the proliferation of IMQ-induced keratinocytes. The treatment attenuated the infiltration of PCNA+ and CD3+ cells in the lesions. Matrine reduced the expression of the MyD88 protein and the inflammatory factors' mRNA in lesional skin, but also in BMDCs (bone marrow derived dendritic cells). These results indicated that matrine suppressed expression of the inflammatory factors by decreasing the expression of the MyD88 protein on the surface of BMDCs, finally alleviating psoriasiform skin lesions. Therefore, the findings suggest that matrine might be a potential candidate for treating psoriasis.
RESUMO
To investigate the apoptosis-induction effect of brucine on human chronic myeloid leukemia cell line K562 cells, K562 cells were exposed to various dosages of brucine. MTT method was used to assayed the growth inhibition effect of brucine on K562 cells. The apoptosis of K562 cells was detected by acridine orange/ethidium bromide (AO/EB) double staining, Annexin-V/PI double labeling method and DNA agarose gel electrophoresis. The results showed that brucine could remarkably inhibit the K562 cell growth in a concentration-dependent and time-dependent manners at the range of 50 to 400 µg/ml, and its most significant inhibition was observed at 400 µg/ml for 72 hours and the inhibition rate was 94.0%. Staining of cells with AO-EB revealed that brucine induced nuclear chromatin condensation. After the K562 cells were treated with the brucine of 400 µg/ml for 72 hours, the most of the nucleus were orange stained and condensation-like or bead-like showing apoptotic morphology. The K562 cells treated with brucine of different concentrations (50, 100, 200, 400, 800 µg/ml) for 72 hours, Annexin-V/PI detection showed brucine could induce apoptosis of K562 cells, and apoptosis rate increased gradually with increasing concentration of drugs. The K562 cells treated with brucine of 400 µg/ml for 72 hours displayed typical ladder strap in DNA gel electrophoresis. It is concluded that brucine can efficiently inhibit cell growth and induce apoptosis of K562 cells with dose-dependent manner in concentrations of 50 - 400 µg/ml.
Assuntos
Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Estricnina/análogos & derivados , Humanos , Células K562 , Estricnina/farmacologiaRESUMO
The aim of this study was to investigate the effect of brucine on secretion of TNF-α, IFN-γ, IL-4 and proliferation of T lymphocytes in patients with aplastic anemia (AA), and to explore its mechanism. Peripheral blood T lymphocytes from 10 patients with AA and 10 healthy volunteers were isolated, purified and cultured. T lymphocytes from the patients were divided into 0, 100, 200 and 400 µg/ml brucine-treated groups. T lymphocytes from healthy volunteers were used as control group. After being cultured for 72 hours, the levels of TNF-α, IFN-γ, IL-4 in the supernatant of cultured T lymphocytes from AA patients were detected by ELISA, and the proliferation of T lymphocytes from AA patients was detected by MTT. The results showed that compared with the normal control group, the levels of TNF-α and IFN-γ in the culture supernatant significantly increased, and IL-4 was significantly decreased. The levels of TNF-α and IFN-γ in the culture supernatant of brucine treated groups were lower, and were dependent on the concentration of brucine. However, the levels of IL-4 were found to be not obviously changed. The inhibition rate of T lymphocytes in 100, 200 and 400µg/ml brucine-treated groups were (13.61 ± 4.31)%, (14.28 ± 4.31)% and (15.12 ± 4.56)% respectively, among which the differences were not statistically significant. It is concluded that the brucine can reduce the levels of TNF-α and IFN-γ through inhibiting the proliferation of T lymphocytes in AA patients, which provides experimental basis for therapy of AA patients.