RESUMO
BACKGROUND: Chronic kidney disease (CKD) is a kidney disease in which there is gradual loss of kidney function over time and end-stage renal disease (ESRD) is the final stage of CKD. Both CKD and ESRD are worldwide health problems with a high economic cost to health systems. However, the molecular mechanisms of the development of CKD and ESRD remain poorly understood. This study aimed to systematically elucidate the molecular mechanisms of the development of CKD and ESRD. METHODS: Transcriptome data of CKD and ESRD were downloaded from the NCBI-GEO database. Differentially expressed genes between cases and controls (chronic kidney disease patients vs. controls, end-stage renal disease patients vs. controls) were calculated using the empirical Bayes algorithm. Gene set enrichment analysis (GSEA) was used for analyzing the KEGG pathway difference between cases and controls. Furthermore, CKD and ESRD target genes were obtained from the Thomson Reuters Integrity database. Tissue-specific gene interaction network analysis was performed using the GIANT web server. RESULTS: There were multiple damaged pathways in ESRD but only a few pathways were disturbed in CKD. Furthermore, we identified 9 dysregulated anti-ESRD genes but no dysregulated anti-CKD genes. Network analysis revealed that the NF-kB signaling pathway was essential for ESRD. CONCLUSIONS: This study revealed several crucial anti-ESRD genes that are involved in the regulation of the NF-kB signaling pathway. This information may be helpful for the treatment of ESRD.
Assuntos
Falência Renal Crônica , Insuficiência Renal Crônica , Teorema de Bayes , Humanos , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/genética , Insuficiência Renal Crônica/genética , TranscriptomaRESUMO
This study is to explore the molecular mechanism of benign bile duct hypertrophic scar formation.Differential proteins between the normal fibroblast (NFB) and scar fibroblast (SCFB) were screened by protein chip assay, and analyzed by pathway-enrichment analysis and function-enrichment analysis. The differential proteins were further tested by ELISA. SiRNA-Act B was transfected to SCFB to down-regulate the expression of Act B. NFB was incubated with rh-Act B. The cell apoptosis and cell cycle were determined by flow cytometry. The expression of Act B, Smad2/3, transforming growth factor-ß1 (TGF-ß1), endothelin-1 (ET-1), thrombospondin-1 (Tsp-1), and Oncostatin M (OSM) were detected by Western blot.A total of 37 differential proteins were identified in SCFBs by microarray (Pâ<â.05), including 27 up-regulated proteins and 10 down-regulated proteins (Pâ<â.05). Their function were associated with Activin signaling, synthesis and degradation of extracellular matrix, formation and activation of cytokine, inflammatory reaction, immunoreaction, tissue damage reaction, cell cycle, migration, apoptosis, and secretion, etc. ELISA results showed that the expression of Act B, TGF-ß1, ET-1 were higher in SCFBs, while the expression of Tsp-1 and OSM were lower in SCFBs (Pâ<â.05). After interfered by siRNA-Act B, the expression of Act B mRNA decreased (Pâ<â.05). The percentage of early apoptosis increased (Pâ<â.05). The expression of Act B, Smad2/3, TGF-ß1 were decreased and Tsp-1, OSM were increased (Pâ<â.05). After treatment with rh-Act B, the percentage of G0/G1 phase of NFBs was decreased and that of S phase was increased without significance (Pâ>â.05). The expression of Act B, Smad2/3, TGF-ß1 were increased (Pâ<â.05) and Tsp-1, OSM were decreased (Pâ<â.01).There are differentially expressed proteins between SCFBs and NFBs. Activin B signal plays an important role in the process of NFB transforming to SCFB, and TGF-ß1, Smad2/3, Tsp-1, and OSM are important participants.
Assuntos
Ativinas/metabolismo , Ductos Biliares/patologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Transdução de Sinais/fisiologia , Adulto , Apoptose/fisiologia , Ciclo Celular/fisiologia , Cicatriz Hipertrófica , Endotelina-1/metabolismo , Matriz Extracelular/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oncostatina M/metabolismo , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Proteína Smad2/metabolismo , Trombospondina 1/metabolismo , Fator de Crescimento Transformador beta1/metabolismoRESUMO
BACKGROUND Worldwide, hepatocellular carcinoma (HCC) accounts for 80-90% of all cases of primary liver cancer, and is one of the ten most common malignancies. This study used bioinformatics analysis to identify genes associated with patient outcome in stages I-IV HCC and the gene pathways that distinguished between normal liver and liver cells and HCC and human HCC cell lines. MATERIAL AND METHODS Target genes were defined as those that had marketed drugs or drugs under development targeting a specific gene and acquired from the Clarivate Analytics Integrity Database. Differential expression gene analysis, co-expression network analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, survival analysis and receiver operating characteristic (ROC) curve analysis were used to explore the similarities and differences in gene expression profiles, functional associations, and survival in stage I-IV HCC. Normal liver cells (HL-7702) and HCC cell lines (HepaRG, HepG2, SK-Hep1, and Huh7) were studied using Western blot and quantitative reverse transcription PCR (RT-qPCR). RESULTS Hierarchical gene clustering identified target genes that distinguished between HCC and normal liver tissue. For stages I-IV HCC, there were seven commonly upregulated target genes EPHB1, LTK, NTRK2, PTK7, TBK1, TIE1, and TLR3, which were mainly involved in immune and signaling transduction pathways. PTK7 was highly expressed in stage I-IV HCC and was an independent prognostic marker for reduced overall survival (OS). CONCLUSIONS Bioinformatics analysis, combined with patient survival analysis, identified PTK7 gene expression as a potential therapeutic target and prognostic biomarker for all stages of HCC.
Assuntos
Carcinoma Hepatocelular/genética , Moléculas de Adesão Celular/genética , Biologia Computacional/métodos , Receptores Proteína Tirosina Quinases/genética , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/metabolismo , Moléculas de Adesão Celular/fisiologia , Linhagem Celular Tumoral , China , Análise por Conglomerados , Bases de Dados Genéticas , Perfilação da Expressão Gênica/métodos , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Estadiamento de Neoplasias , Prognóstico , Mapas de Interação de Proteínas/genética , Curva ROC , Receptores Proteína Tirosina Quinases/fisiologia , Transcriptoma/genéticaRESUMO
Melanoma is usually driven by mutations in BRAF or NRAS, which trigger hyperactivation of MAPK signaling. However, MAPK-targeted therapies are not sustainably effective in most patients. Accordingly, characterizing mechanisms that co-operatively drive melanoma progression is key to improving patient outcomes. One possible mechanism is the Hippo signaling pathway, which regulates cancer progression via its central oncoproteins YAP and TAZ, although is thought to be only rarely affected by direct mutation. As YAP hyperactivation occurs in uveal melanoma, we investigated this oncogene in cutaneous melanoma. YAP protein expression was elevated in most benign nevi and primary cutaneous melanomas but present at only very low levels in normal melanocytes. In patient-derived xenografts and melanoma cell lines, we observed variable reliance of cell viability on Hippo pathway signaling that was independent of TAZ activity and also of classical melanoma driver mutations such as BRAF and NRAS. Finally, in genotyping studies of melanoma, we observed the first ever hyperactivating YAP mutations in a human cancer, manifest as seven distinct missense point mutations that caused serine to alanine transpositions. Strikingly, these mutate four serine residues known to be targeted by the Hippo pathway and we show that they lead to hyperactivation of YAP. IMPLICATIONS: Our studies highlight the YAP oncoprotein as a potential therapeutic target in select subgroups of melanoma patients, although successful treatment with anti-YAP therapies will depend on identification of biomarkers additional to YAP protein expression.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Biomarcadores Tumorais/genética , Sequenciamento do Exoma , Melanoma/genética , Neoplasias Cutâneas/genética , Fatores de Transcrição/genética , Aciltransferases , Animais , Linhagem Celular Tumoral , Feminino , GTP Fosfo-Hidrolases/genética , Regulação Neoplásica da Expressão Gênica/genética , Genótipo , Via de Sinalização Hippo , Humanos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Melanoma/patologia , Proteínas de Membrana/genética , Camundongos , Mutação/genética , Células NIH 3T3 , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas B-raf/genética , Transdução de Sinais/genética , Neoplasias Cutâneas/patologia , Ensaios Antitumorais Modelo de Xenoenxerto , Proteínas de Sinalização YAP , Melanoma Maligno CutâneoRESUMO
This study aims to explore the effect of PKC-α gene silencing on the occurrence of ultrafiltration failure (UFF) in peritoneal dialysis (PD) rats. Forty-eight male SD rats were collected to establish 5/6 renal resection uremic and uremic PD rats models. Rats were assigned into control, sham operation, uremia, PD-2 W (peritoneal dialysis for 2 weeks), PD-4 W (peritoneal dialysis for 4 weeks), negative control (NC) (peritoneal dialysis for 4 weeks, and injected 0.1 mg/kg blank plasmid into abdominal cavity) and PKC-α siRNA (peritoneal dialysis for 4 weeks, and injected 0.1 mg/kg PKC-α siRNA into abdominal cavity) groups. CD34 staining was performed to determine microvessel density (MVD) for peritoneal tissues. The mRNA and protein expression of PKC-α in peritoneal tissue were detected by qRT-PCR and Western blot. Compared with the control group, MVD, the mRNA and protein expression of PKC-α were significantly increased in rats of the uremia, PD-2 W, PD-4 W, NC, and PKC-α siRNA groups. Compared with the uremia group, MVD, the mRNA and protein expression of PKC-α were increased, the changes observed in the PD-4 W and NC groups were better obvious than in the PD-2 W group. In comparison with the PD-4 W and NC groups, MVD, the mRNA and protein expression of PKC-α in rats were decreased in the PKC-α siRNA group. PKC-α gene has a high expression in uremic PD rats, and PKC-α gene silencing is able to increase UF while decrease MVD and glucose transport in peritoneal tissues thus reversing UFF in PD rats. J. Cell. Biochem. 118: 4607-4616, 2017. © 2017 Wiley Periodicals, Inc.
Assuntos
Inativação Gênica , Hemodiafiltração , Diálise Peritoneal , Proteína Quinase C-alfa , RNA Interferente Pequeno , Animais , Modelos Animais de Doenças , Masculino , Proteína Quinase C-alfa/antagonistas & inibidores , Proteína Quinase C-alfa/genética , Proteína Quinase C-alfa/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Ratos , Ratos Sprague-Dawley , Uremia/enzimologia , Uremia/genética , Uremia/patologia , Uremia/terapiaRESUMO
Aberrant microRNA (miRNA or miR) expression has been reported to contribute to the pathogenesis of hepatocellular carcinoma (HCC). However, the involvement of specific miRNAs in HCC remains to be elucidated. The present study aimed to investigate the potential role of miR-200b and the mechanism underlying its function in hepatocarcinogenesis. The results of the present study demonstrated that the expression levels of miR200b were significantly reduced in HCC tissue samples, as compared with normal liver (NL) and paratumorous (PT) tissue samples. The results also revealed that miR200b expression levels in HepG2 cells were significantly decreased compared with those in L02 cells. In addition, western blotting and reverse transcriptionquantitative polymerase chain reaction demonstrated that the expression levels of DNA methyltransferase 3a (DNMT3a), a possible target gene for miR200b, were significantly higher in HCC tissue samples, as compared with those in NL and PT tissue samples. Furthermore, the data suggested that DNMT3a was a direct target gene of miR200b. Upregulated miR200b expression in HepG2 cells led to a decrease in DNMT3a expression levels, and an inhibition of cell proliferation. These results suggested that miR200b has an important role in hepatocarcinogenesis and acts by downregulating DNMT3a expression. Thus, miR-200b may be a promising target for HCC treatment.
Assuntos
Carcinoma Hepatocelular/metabolismo , Proliferação de Células , DNA (Citosina-5-)-Metiltransferases/biossíntese , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/metabolismo , MicroRNAs/biossíntese , Proteínas de Neoplasias/biossíntese , RNA Neoplásico/biossíntese , Adulto , Idoso , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , DNA (Citosina-5-)-Metiltransferases/genética , DNA Metiltransferase 3A , Feminino , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , RNA Neoplásico/genéticaRESUMO
OBJECTIVE: The angiotensin-converting enzyme gene is one of the most studied candidate genes related to atrial fibrillation. Among the polymorphisms of the angiotensin-converting enzyme gene, the 2350 G/A polymorphism (rs4343) is known to have the most significant effects on the plasma angiotensin-converting enzyme concentration. The aim of the present study was to investigate the association of the angiotensin-converting enzyme 2350 G/A polymorphism with atrial fibrillation in Han Chinese patients with essential hypertension. METHODS: A total of 169 hypertensive patients were eligible for this study. Patients with atrial fibrillation (nâ=â75) were allocated to the atrial fibrillation group, and 94 subjects without atrial fibrillation were allocated to the control group. The PCR-based restriction fragment length polymorphism technique was used to assess the genotype frequencies. RESULTS: The distributions of the angiotensin-converting enzyme 2350 G/A genotypes (GG, GA, and AA, respectively) were 40.43%, 41.49%, and 18.08% in the controls and 18.67%, 46.67%, and 34.66% in the atrial fibrillation subjects (pâ=â0.037). The frequency of the A allele in the atrial fibrillation group was significantly greater than in the control group (58.00% vs. 38.83%, pâ=â0.0007). Compared with the wild-type GG genotype, the GA and AA genotypes had an increased risk for atrial fibrillation. Additionally, atrial fibrillation patients with the AA genotype had greater left atrial dimensions than the patients with the GG or GA genotypes (p<0.01 and p<0.05, respectively). CONCLUSIONS: The results obtained in this study indicate that the angiotensin-converting enzyme 2350 G/A polymorphism is associated with atrial fibrillation and that the A allele shows an increased risk for atrial fibrillation in Han Chinese patients with essential hypertension.
Assuntos
Fibrilação Atrial/genética , Hipertensão/genética , Peptidil Dipeptidase A/genética , Polimorfismo Genético/genética , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Povo Asiático/genética , Pressão Sanguínea/genética , Índice de Massa Corporal , China , Hipertensão Essencial , Feminino , Frequência do Gene , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Fatores de Risco , Fatores SexuaisRESUMO
OBJECTIVE: The angiotensin-converting enzyme gene is one of the most studied candidate genes related to atrial fibrillation. Among the polymorphisms of the angiotensin-converting enzyme gene, the 2350 G/A polymorphism (rs4343) is known to have the most significant effects on the plasma angiotensin-converting enzyme concentration. The aim of the present study was to investigate the association of the angiotensin-converting enzyme 2350 G/A polymorphism with atrial fibrillation in Han Chinese patients with essential hypertension. METHODS: A total of 169 hypertensive patients were eligible for this study. Patients with atrial fibrillation (n = 75) were allocated to the atrial fibrillation group, and 94 subjects without atrial fibrillation were allocated to the control group. The PCR-based restriction fragment length polymorphism technique was used to assess the genotype frequencies. RESULTS: The distributions of the angiotensin-converting enzyme 2350 G/A genotypes (GG, GA, and AA, respectively) were 40.43%, 41.49%, and 18.08% in the controls and 18.67%, 46.67%, and 34.66% in the atrial fibrillation subjects (p = 0.037). The frequency of the A allele in the atrial fibrillation group was significantly greater than in the control group (58.00% vs. 38.83%, p = 0.0007). Compared with the wild-type GG genotype, the GA and AA genotypes had an increased risk for atrial fibrillation. Additionally, atrial fibrillation patients with the AA genotype had greater left atrial dimensions than the patients with the GG or GA genotypes (p<0.01 and p<0.05, respectively). CONCLUSIONS: The results obtained in this study indicate that the angiotensin-converting enzyme 2350 G/A polymorphism is associated with atrial fibrillation and that the A allele shows an increased risk for atrial fibrillation in Han Chinese patients with essential hypertension. .
Assuntos
Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fibrilação Atrial/genética , Hipertensão/genética , Peptidil Dipeptidase A/genética , Polimorfismo Genético/genética , Fatores Etários , Povo Asiático/genética , Índice de Massa Corporal , Pressão Sanguínea/genética , China , Frequência do Gene , Predisposição Genética para Doença , Reação em Cadeia da Polimerase , Fatores de Risco , Fatores SexuaisRESUMO
To define genetic lesions driving leukemia, we targeted cre-dependent Sleeping Beauty (SB) transposon mutagenesis to the blood-forming system using a hematopoietic-selective vav 1 oncogene (vav1) promoter. Leukemias of diverse lineages ensued, most commonly lymphoid leukemia and erythroleukemia. The inclusion of a transgenic allele of Janus kinase 2 (JAK2)V617F resulted in acceleration of transposon-driven disease and strong selection for erythroleukemic pathology with transformation of bipotential erythro-megakaryocytic cells. The genes encoding the E-twenty-six (ETS) transcription factors Ets related gene (Erg) and Ets1 were the most common sites for transposon insertion in SB-induced JAK2V617F-positive erythroleukemias, present in 87.5% and 65%, respectively, of independent leukemias examined. The role of activated Erg was validated by reproducing erythroleukemic pathology in mice transplanted with fetal liver cells expressing translocated in liposarcoma (TLS)-ERG, an activated form of ERG found in human leukemia. Via application of SB mutagenesis to TLS-ERG-induced erythroid transformation, we identified multiple loci as likely collaborators with activation of Erg. Jak2 was identified as a common transposon insertion site in TLS-ERG-induced disease, strongly validating the cooperation between JAK2V617F and transposon insertion at the Erg locus in the JAK2V617F-positive leukemias. Moreover, loci expressing other regulators of signal transduction pathways were conspicuous among the common transposon insertion sites in TLS-ERG-driven leukemia, suggesting that a key mechanism in erythroleukemia may be the collaboration of lesions disturbing erythroid maturation, most notably in genes of the ETS family, with mutations that reduce dependence on exogenous signals.
Assuntos
Regulação Leucêmica da Expressão Gênica , Janus Quinase 2/metabolismo , Leucemia Eritroblástica Aguda/metabolismo , Leucemia Megacarioblástica Aguda/metabolismo , Proteínas Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-ets/metabolismo , Alelos , Animais , Elementos de DNA Transponíveis , Genótipo , Janus Quinase 2/genética , Leucemia Eritroblástica Aguda/genética , Leucemia Megacarioblástica Aguda/genética , Camundongos , Camundongos Transgênicos , Mutagênese , Transplante de Neoplasias , Proteínas Oncogênicas/genética , Regiões Promotoras Genéticas , Proteína Proto-Oncogênica c-ets-1/genética , Proteína Proto-Oncogênica c-ets-1/metabolismo , Proteínas Proto-Oncogênicas c-ets/genética , Recombinação Genética , Análise de Sequência de DNA , Transdução de Sinais/genética , Fatores de Transcrição , Regulador Transcricional ERGRESUMO
AIM: To construct the recombinant adenoviral vector which can efficiently inhibit mature miR-21 expression and explore its effects and the underlying mechanisms on hepatoma cell line HepG2. METHODS: Based on miRNA-sponge technology, we synthesized 8 duplicated fragments, fully complementary with the miR-21 sequence and cloned them into a shuttle vector pAdTrack-CMV. The constructed plasmid was sequenced and linearized for homologous recombination with pAdEasy-1 vector in BJ5183 bacteria, then transfected into 293A cells. The recombinant adenovirus was used to challenge HepG2 cells. Mature miR-21 level was detected by real-time PCR. Apoptosis and proliferation of the HepG2 cells were detected by Hochest 33258 staining, Western blotting and MTT assay. RESULTS: The restriction enzyme digestion, DNA sequencing and detection of GFP expression demonstrated that recombinant adenoviral vector was constructed successfully. The recombinant adenovirus inhibited the expression of miR-21 in HepG2 cells, and also depressed the proliferation of HepG2 cells and promoted the apoptosis. CONCLUSION: The recombinant adenoviral vector we successfully constructed could efficiently reduce the expression of miR-21 in HepG2 cells and depress the proliferation of HepG2 cells.
Assuntos
Adenoviridae/fisiologia , Vetores Genéticos/fisiologia , Neoplasias Hepáticas/genética , MicroRNAs/genética , Montagem de Vírus , Adenoviridae/genética , Apoptose , Sequência de Bases , Proliferação de Células , Regulação para Baixo , Vetores Genéticos/genética , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/fisiopatologia , Neoplasias Hepáticas/terapia , MicroRNAs/metabolismo , MicroRNAs/uso terapêutico , Dados de Sequência MolecularRESUMO
OBJECTIVES: To determine the effect of 5-aza-2'-deoxycytidine (DAC) on human endometrial carcinoma cell (HECC) oncogenicity and demonstrate a molecular mechanism by which DAC modulates HECC oncogenicity. METHODS: The effect of DAC was tested on HECC RL95-2, AN3, Ishikawa and ECC1 cells. The role of 14-3-3σ on HECC oncogenicity in response to DAC treatment was evaluated in RL95-2 and AN3 cells after forced expression or silencing of 14-3-3σ gene expression. RESULTS: Treatment of HECC with DAC produced non-cytotoxic cell growth inhibition and G2/M cell cycle arrest. This effect was strongly correlated with increased expression of p21 and 14-3-3σ. Silencing of 14-3-3σ induced cellular proliferation and reduced the effect of DAC on cell cycle arrest in G2/M phases. Conversely, forced expression of 14-3-3σ showed the opposite effect. Furthermore, forced expression of 14-3-3σ in human endometrial cell lines reduced cell growth and colony formation. CONCLUSIONS: We suggest that 14-3-3σ in HECC suppresses cell proliferation and mediates DAC induced G2/M arrest and inhibition of cell proliferation in HECC.
Assuntos
Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Azacitidina/análogos & derivados , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Exonucleases/genética , Exonucleases/metabolismo , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase M do Ciclo Celular/efeitos dos fármacos , Proteínas 14-3-3/biossíntese , Proteínas 14-3-3/deficiência , Azacitidina/farmacologia , Biomarcadores Tumorais/biossíntese , Biomarcadores Tumorais/deficiência , Processos de Crescimento Celular/efeitos dos fármacos , Processos de Crescimento Celular/genética , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Metilação de DNA/efeitos dos fármacos , Decitabina , Neoplasias do Endométrio/tratamento farmacológico , Neoplasias do Endométrio/metabolismo , Exonucleases/biossíntese , Exonucleases/deficiência , Exorribonucleases , Feminino , Pontos de Checagem da Fase G2 do Ciclo Celular/genética , Genes Supressores de Tumor/efeitos dos fármacos , Humanos , Pontos de Checagem da Fase M do Ciclo Celular/genética , TransfecçãoRESUMO
We herein demonstrate an oncogenic role for signal transducer and activator of transcription (STAT)-3alpha (the full length STAT3 isoform), which also mediates autocrine human GH (hGH)-stimulated oncogenicity, in human endometrial carcinoma (EC) cells. Autocrine hGH stimulated Y705 phosphorylation of STAT3 and STAT3-mediated transcriptional activity in a SRC and Janus-2 Kinase dependent manner in human EC cell lines. Forced expression of a constitutively active variant of STAT3alpha increased proliferation, anchorage-independent, three-dimensional (3D) Matrigel, and xenograft growth and promoted epithelial-mesenchymal transition, migration, and invasion of EC cells. Conversely, the oncogenic capacity of EC cells was significantly impaired by treatment with JSI-124, an inhibitor of STAT3 phosphorylation and activity, small interfering RNA-mediated depletion of STAT3alpha, or a dominant-negative variant of STAT3alpha. Furthermore, the enhanced EC cell oncogenicity stimulated by autocrine hGH, was also abrogated by functional inhibition or small interfering RNA-mediated depletion of STAT3alpha. STAT3alpha may therefore be a common mediator of oncogenic signaling pathways stimulating progression of EC.
Assuntos
Neoplasias do Endométrio/metabolismo , Hormônio do Crescimento Humano/metabolismo , Proteínas de Membrana/metabolismo , Fator de Transcrição STAT3/metabolismo , Animais , Comunicação Autócrina/efeitos dos fármacos , Western Blotting , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Feminino , Hormônio do Crescimento Humano/genética , Hormônio do Crescimento Humano/farmacologia , Humanos , Proteínas de Membrana/genética , Camundongos , Camundongos SCID , Microscopia de Fluorescência , Invasividade Neoplásica , Neoplasias Experimentais/genética , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Fosforilação , Interferência de RNA , Fator de Transcrição STAT3/genética , Transplante Heterólogo , Triterpenos/farmacologiaRESUMO
We herein show that Artemin (ARTN), one of the glial cell line-derived neurotrophic factor family of ligands, promotes progression of human non-small cell lung carcinoma (NSCLC). Oncomine data indicate that expression of components of the ARTN signaling pathway (ARTN, GFRA3, and RET) is increased in neoplastic compared with normal lung tissues; increased expression of ARTN in NSCLC also predicted metastasis to lymph nodes and a higher grade in certain NSCLC subtypes. Forced expression of ARTN stimulated survival, anchorage-independent, and three-dimensional Matrigel growth of NSCLC cell lines. ARTN increased BCL2 expression by transcriptional upregulation, and inhibition of BCL2 abrogated the oncogenic properties of ARTN in NSCLC cells. Forced expression of ARTN also enhanced migration and invasion of NSCLC cells. Forced expression of ARTN in H1299 cells additionally resulted in larger xenograft tumors, which were highly proliferative, invasive, and metastatic. Concordantly, either small interfering RNA-mediated depletion or functional inhibition of endogenous ARTN with antibodies reduced oncogenicity and invasiveness of NSCLC cells. ARTN therefore mediates progression of NSCLC and may be a potential therapeutic target for NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Progressão da Doença , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Proteínas do Tecido Nervoso/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Carcinoma Pulmonar de Células não Pequenas/genética , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Colágeno/metabolismo , Combinação de Medicamentos , Regulação Neoplásica da Expressão Gênica , Humanos , Laminina/metabolismo , Neoplasias Pulmonares/genética , Camundongos , Invasividade Neoplásica , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Proteínas do Tecido Nervoso/genética , Proteoglicanas/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Increased activation of signal transducer and activator of transcription (STAT)-5 has been reported in various malignancies including mammary carcinoma. However, it is only recently that potentially distinct roles of STAT5A and STAT5B in neoplasia have begun to emerge. Herein we systematically delineate the functions of STAT5A and STAT5B in human mammary carcinoma cell lines MCF-7 and T47D. Forced expression of constitutively active (CA) STAT5A enhanced both survival and anchorage-independent growth of human mammary carcinoma cells but concordantly suppressed cell motility as revealed in colony scattering, cell migration, and invasion assays. In contrast, forced expression of CA STAT5B exhibited lower potency than CA STAT5A in enhancing survival and anchorage-independent growth of mammary carcinoma cells and exerted no effects on cell motility. Differential expression of genes that regulate cellular survival and motility was concomitantly observed on forced expression of CA STAT5A or CA STAT5B. Small interfering RNA-mediated depletion of STAT5A significantly impaired anchorage-independent growth of human mammary carcinoma cells, whereas a smaller reduction was observed upon small interfering RNA-mediated depletion of STAT5B. Depletion of endogenous STAT5A also significantly enhanced cell motility, whereas depletion of endogenous STAT5B exhibited no effect. Xenograft studies provided data concordant with the in vitro effects of the two STAT5 isoforms. We therefore demonstrate that STAT5A and STAT5B differentially regulate behavior of human mammary carcinoma cells.
Assuntos
Neoplasias da Mama/patologia , Carcinoma/patologia , Movimento Celular , Proliferação de Células , Fator de Transcrição STAT5/fisiologia , Proteínas Supressoras de Tumor/fisiologia , Animais , Neoplasias da Mama/genética , Carcinoma/genética , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Camundongos Nus , RNA Interferente Pequeno/farmacologia , Fator de Transcrição STAT5/antagonistas & inibidores , Fator de Transcrição STAT5/genética , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor/antagonistas & inibidores , Proteínas Supressoras de Tumor/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: To study the protective effect of reduced glutathione (GSH) on multiple organ function in patient with acute pancreatitis. METHODS: A total of 45 patients with acute pancreatitis was randomly divided into two groups, 22 patients were given GSH in a dose of 1.2 g/d through intravenous drip for 7 days as GSH group, and 23 patients were not given GSH to serve as control group. The same treatment was given to both groups other than GSH. Plasma contents of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) and biochemistry indexes were determined. RESULTS: After the treatment, the levels of TNF-alpha and IL-6 were significantly decreased in both groups (all P<0.05). The levels of TNF-alpha and IL-6 were decreased much more obviously in GSH group compared with those in control group (P<0.05). Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (Cr), blood urea nitrogen (BUN), lactate dehydrogenase (LDH), creatine kinase (CK), and MB isoenzyme of creatine kinase (CK-MB) were significantly decreased in both treatment groups, but the decrease of all these indexes was more marked in GSH group (P<0.05). CONCLUSION: The results suggest that GSH has beneficial effects in protecting visceral organ function in patients with acute pancreatitis.