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1.
Life Sci ; 244: 117300, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31953162

RESUMO

MiR-346-5p is overexpressed in several cancers, including colorectal cancer (CRC). However, the effects of miR-346-5p on CRC progression have not yet been clarified. In our study, miR-346-5p levels in four CRC cell lines and normal human colon epithelial cells were determined by real-time PCR. SW620 and HCT116 cells were selected and then transfected with miR-346-5p mimic, miR-346-5p inhibitor, or specific siRNAs targeting F-box/LRR-repeat protein 2 (FBXL2). Cell proliferation, cell cycle distribution and cell cycle regulators were examined by CCK-8 assay, flow cytometry, and western blot. The binding of miR-346-5p on 3' untranslated region (UTR) of FBXL2 were verified by dual-luciferase reporter assay. CRC cells were co-transfected with miR-346-5p inhibitor and siFBXL2 to investigate the involvement of FBXL2. Interaction of FBXL2 with forkhead box M1 (FoxM1) was examined by co-immunoprecipitation (Co-IP) assay. The effect of miR-346-5p knockdown on CRC tumorigenesis in vivo was investigated. Here, we found that miR-346-5p overexpression promoted, while miR-346-5p knockdown inhibited cell proliferation and G1-S transition. Inhibition of FBXL2 showed similar effects as miR-346-5p overexpression. Moreover, we verified that FBXL2 was a direct target of miR-346-5p. FBXL2 interacted with FoxM1, and then negatively regulated both FoxM1 and nuclear ß-catenin levels. Additionally, FBXL2 knockdown reversed the effects of miR-346-5p inhibitor. In xenograft models, miR-346-5p knockdown significantly inhibited tumor growth, increased FBXL2 expression, and downregulated the levels of FoxM1 and nuclear ß-catenin. In conclusion, miR-346-5p may promote CRC growth by targeting FBXL2 and activating the ß-catenin signaling pathway. MiR-346-5p may be a novel target in cancer therapy.


Assuntos
Neoplasias Colorretais/metabolismo , Proteínas F-Box/metabolismo , MicroRNAs/genética , Animais , Apoptose/genética , Carcinogênese/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Colorretais/genética , Proteínas F-Box/genética , Proteína Forkhead Box M1/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , Transdução de Sinais/genética , Via de Sinalização Wnt/genética , Ensaios Antitumorais Modelo de Xenoenxerto , beta Catenina/metabolismo
2.
DNA Cell Biol ; 39(1): 118-125, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31859543

RESUMO

Colorectal cancer (CRC) has been suggested to be one of the leading cancer types all over the world. Till now, the molecular mechanism by which circCCT3 regulates CRC remains to be clarified. To detect mRNA and protein levels of various genes, Reverse Transcription-quantitative PCR and western blot were used in our study. Luciferase reporter assay was utilized to probe direct interaction between genes. We used transwell assay to assess the invasion ability of CRC cells. For apoptosis detection, immunofluorescence of CRC cells by Annexin V staining was performed. We carried out bioinformatic analysis to show higher expression of circCCT3 in human clinical CRC tumors. Low level of circCCT3 was closely associated with higher disease-free survival of CRC patients. Moreover, we found that circCCT3 was linked to advanced stage of CRC. miR-613 is the target of circCCT3 and responsible for circCCT3-modulated invasion and apoptosis of CRC cells. In addition, we identified WNT3 and vascular endothelial growth factor A (VEGFA) as downstream effectors of miR-613 in CRC cells. WNT3 and VEGFA overexpression resulted in partial rescue of miR-613-mediated phenotypes of CRC cells. In conclusion, we propose that circCCT3 contributes to CRC metastasis via miR-613/WNT3 or miR-613/VEGFA, promoting the development of therapeutical approaches for treating CRC.


Assuntos
Chaperonina com TCP-1/genética , Neoplasias Colorretais/genética , MicroRNAs/genética , Fator A de Crescimento do Endotélio Vascular/genética , Via de Sinalização Wnt/genética , Regiões 3' não Traduzidas/genética , Sequência de Bases , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica , Células HCT116 , Humanos , Estimativa de Kaplan-Meier , Metástase Neoplásica , Homologia de Sequência do Ácido Nucleico , Proteína Wnt3/genética
3.
Nat Genet ; 51(12): 1723-1731, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31784729

RESUMO

WNT signaling activates MYC expression in cancer cells. Here we report that this involves an oncogenic super-enhancer-mediated tethering of active MYC alleles to nuclear pores to increase transcript export rates. As the decay of MYC transcripts is more rapid in the nucleus than in the cytoplasm, the oncogenic super-enhancer-facilitated export of nuclear MYC transcripts expedites their escape from the nuclear degradation system in colon cancer cells. The net sum of this process, as supported by computer modeling, is greater cytoplasmic MYC messenger RNA levels in colon cancer cells than in wild type cells. The cancer-cell-specific gating of MYC is regulated by AHCTF1 (also known as ELYS), which connects nucleoporins to the oncogenic super-enhancer via ß-catenin. We conclude that WNT signaling collaborates with chromatin architecture to post-transcriptionally dysregulate the expression of a canonical cancer driver.


Assuntos
Proteínas de Ligação a DNA/genética , Elementos Facilitadores Genéticos , Genes myc , Fatores de Transcrição/genética , Via de Sinalização Wnt/genética , Colo/citologia , Proteínas de Ligação a DNA/metabolismo , Células Epiteliais/fisiologia , Regulação Neoplásica da Expressão Gênica , Células HCT116 , Humanos , Antígenos de Histocompatibilidade Menor/genética , Antígenos de Histocompatibilidade Menor/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Transporte Proteico , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Proto-Oncogênicas c-myc/metabolismo , Processamento Pós-Transcricional do RNA , Fatores de Transcrição/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
4.
Postepy Biochem ; 65(3): 183-192, 2019 10 01.
Artigo em Polonês | MEDLINE | ID: mdl-31643165

RESUMO

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


Assuntos
Via de Sinalização Wnt , beta Catenina/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Humanos , Neoplasias/metabolismo , Neoplasias/patologia , Fatores de Transcrição TCF/metabolismo , Via de Sinalização Wnt/genética
5.
PLoS Genet ; 15(10): e1008401, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31626630

RESUMO

Wnt signaling regulates primary body axis formation across the Metazoa, with high Wnt signaling specifying posterior identity. Whether a common Wnt-driven transcriptional program accomplishes this broad role is poorly understood. We identified genes acutely affected after Wnt signaling inhibition in the posterior of two regenerative species, the planarian Schmidtea mediterranea and the acoel Hofstenia miamia, which are separated by >550 million years of evolution. Wnt signaling was found to maintain positional information in muscle and regional gene expression in multiple differentiated cell types. sp5, Hox genes, and Wnt pathway components are down-regulated rapidly after ß-catenin RNAi in both species. Brachyury, a vertebrate Wnt target, also displays Wnt-dependent expression in Hofstenia. sp5 inhibits trunk gene expression in the tail of planarians and acoels, promoting separate tail-trunk body domains. A planarian posterior Hox gene, Post-2d, promotes normal tail regeneration. We propose that common regulation of a small gene set-Hox, sp5, and Brachyury-might underlie the widespread utilization of Wnt signaling in primary axis patterning across the Bilateria.


Assuntos
Padronização Corporal/genética , Genes Homeobox/genética , Planárias/genética , Regeneração/genética , Animais , Diferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Desenvolvimento Muscular/genética , Proteínas Nucleares/genética , Planárias/crescimento & desenvolvimento , Proteínas Wnt/genética , Via de Sinalização Wnt/genética
6.
Nat Commun ; 10(1): 4710, 2019 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-31624259

RESUMO

Morphogen signalling forms an activity gradient and instructs cell identities in a signalling strength-dependent manner to pattern developing tissues. However, developing tissues also undergo dynamic morphogenesis, which may produce cells with unfit morphogen signalling and consequent noisy morphogen gradients. Here we show that a cell competition-related system corrects such noisy morphogen gradients. Zebrafish imaging analyses of the Wnt/ß-catenin signalling gradient, which acts as a morphogen to establish embryonic anterior-posterior patterning, identify that unfit cells with abnormal Wnt/ß-catenin activity spontaneously appear and produce noise in the gradient. Communication between unfit and neighbouring fit cells via cadherin proteins stimulates apoptosis of the unfit cells by activating Smad signalling and reactive oxygen species production. This unfit cell elimination is required for proper Wnt/ß-catenin gradient formation and consequent anterior-posterior patterning. Because this gradient controls patterning not only in the embryo but also in adult tissues, this system may support tissue robustness and disease prevention.


Assuntos
Padronização Corporal/genética , Embrião não Mamífero/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Morfogênese/genética , Via de Sinalização Wnt/genética , Proteínas de Peixe-Zebra/genética , beta Catenina/genética , Animais , Animais Geneticamente Modificados , Apoptose/genética , Embrião não Mamífero/citologia , Embrião não Mamífero/embriologia , Microscopia de Fluorescência , Imagem com Lapso de Tempo/métodos , Peixe-Zebra , Proteínas de Peixe-Zebra/metabolismo , beta Catenina/metabolismo
7.
Nat Commun ; 10(1): 4184, 2019 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-31519875

RESUMO

Axin is a key scaffolding protein responsible for the formation of the ß-catenin destruction complex. Stability of Axin protein is regulated by the ubiquitin-proteasome system, and modulation of cellular concentration of Axin protein has a profound effect on Wnt/ß-catenin signaling. Although E3s promoting Axin ubiquitination have been identified, the deubiquitinase responsible for Axin deubiquitination and stabilization remains unknown. Here, we identify USP7 as a potent negative regulator of Wnt/ß-catenin signaling through CRISPR screens. Genetic ablation or pharmacological inhibition of USP7 robustly increases Wnt/ß-catenin signaling in multiple cellular systems. USP7 directly interacts with Axin through its TRAF domain, and promotes deubiquitination and stabilization of Axin. Inhibition of USP7 regulates osteoblast differentiation and adipocyte differentiation through increasing Wnt/ß-catenin signaling. Our study reveals a critical mechanism that prevents excessive degradation of Axin and identifies USP7 as a target for sensitizing cells to Wnt/ß-catenin signaling.


Assuntos
Proteína Axina/metabolismo , Peptidase 7 Específica de Ubiquitina/metabolismo , beta Catenina/metabolismo , Células 3T3-L1 , Adipócitos/metabolismo , Animais , Proteína Axina/genética , Linhagem Celular , Linhagem Celular Tumoral , Citometria de Fluxo , Células HCT116 , Humanos , Imunoprecipitação , Camundongos , Osteoblastos/metabolismo , Estabilidade Proteica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Peptidase 7 Específica de Ubiquitina/genética , Ubiquitinação/genética , Ubiquitinação/fisiologia , Via de Sinalização Wnt/genética , Via de Sinalização Wnt/fisiologia , beta Catenina/genética
8.
Nat Commun ; 10(1): 4368, 2019 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-31554819

RESUMO

The colonic epithelial turnover is driven by crypt-base stem cells that express the R-spondin receptor Lgr5. Signals that regulate epithelial regeneration upon stem cell injury are largely unknown. Here, we explore the dynamics of Wnt signaling in the colon. We identify two populations of cells with active Wnt signaling: highly proliferative Lgr5+/Axin2+ cells, as well as secretory Lgr5-/Axin2+ cells. Upon Lgr5+ cell depletion, these cells are recruited to contribute to crypt regeneration. Chemical injury induced by DSS leads to a loss of both Lgr5+ cells and Axin2+ cells and epithelial regeneration is driven by Axin2- cells, including differentiated Krt20+ surface enterocytes. Regeneration requires stromal Rspo3, which is present at increased levels upon injury and reprograms Lgr5- but Lgr4+ differentiated cells. In contrast, depletion of stromal Rspo3 impairs crypt regeneration, even upon mild injury. We demonstrate that Rspo3 is essential for epithelial repair via induction of Wnt signaling in differentiated cells.


Assuntos
Colo/fisiologia , Mucosa Intestinal/fisiologia , Regeneração/fisiologia , Células-Tronco/metabolismo , Trombospondinas/metabolismo , Animais , Proteína Axina/genética , Proteína Axina/metabolismo , Diferenciação Celular/genética , Colite/genética , Colite/metabolismo , Colo/metabolismo , Enterócitos/metabolismo , Perfilação da Expressão Gênica/métodos , Mucosa Intestinal/metabolismo , Queratina-20/genética , Queratina-20/metabolismo , Camundongos Knockout , Camundongos Transgênicos , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Regeneração/genética , Células-Tronco/citologia , Trombospondinas/genética , Via de Sinalização Wnt/genética
9.
Nat Commun ; 10(1): 4251, 2019 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-31534175

RESUMO

The paralogous scaffold proteins axin and conductin/axin2 are key factors in the negative regulation of the Wnt pathway transcription factor ß-catenin, thereby representing interesting targets for signaling regulation. Polymerization of axin proteins is essential for their activity in suppressing Wnt/ß-catenin signaling. Notably, conductin shows less polymerization and lower activity than axin. By domain swapping between axin and conductin we here identify an aggregation site in the conductin RGS domain which prevents conductin polymerization. Induction of conductin polymerization by point mutations of this aggregon results in enhanced inhibition of Wnt/ß-catenin signaling. Importantly, we identify a short peptide which induces conductin polymerization via masking the aggregon, thereby enhancing ß-catenin degradation, inhibiting ß-catenin-dependent transcription and repressing growth of colorectal cancer cells. Our study reveals a mechanism for regulating signaling pathways via the polymerization status of scaffold proteins and suggests a strategy for targeted colorectal cancer therapy.


Assuntos
Proteína Axina/metabolismo , Neoplasias Colorretais/patologia , Via de Sinalização Wnt/genética , beta Catenina/metabolismo , Proteína Axina/genética , Sistemas CRISPR-Cas/genética , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias Colorretais/genética , Técnicas de Inativação de Genes , Células HEK293 , Humanos , Proteínas Wnt/metabolismo , Via de Sinalização Wnt/fisiologia
10.
Nat Neurosci ; 22(10): 1624-1634, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31551593

RESUMO

Hundreds of genes are implicated in autism spectrum disorder (ASD), but the mechanisms through which they contribute to ASD pathophysiology remain elusive. Here we analyzed leukocyte transcriptomics from 1- to 4-year-old male toddlers with ASD or typical development from the general population. We discovered a perturbed gene network that includes highly expressed genes during fetal brain development. This network is dysregulated in human induced pluripotent stem cell-derived neuron models of ASD. High-confidence ASD risk genes emerge as upstream regulators of the network, and many risk genes may impact the network by modulating RAS-ERK, PI3K-AKT and WNT-ß-catenin signaling pathways. We found that the degree of dysregulation in this network correlated with the severity of ASD symptoms in the toddlers. These results demonstrate how the heterogeneous genetics of ASD may dysregulate a core network to influence brain development at prenatal and very early postnatal ages and, thereby, the severity of later ASD symptoms.


Assuntos
Transtorno do Espectro Autista/genética , Redes Reguladoras de Genes/genética , Transtorno do Espectro Autista/patologia , Encéfalo/embriologia , Encéfalo/patologia , Pré-Escolar , Desenvolvimento Fetal/genética , Humanos , Lactente , Leucócitos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Mutação/genética , Células-Tronco Neurais , Proteína Oncogênica v-akt/genética , Fosfatidilinositol 3-Quinases/genética , Transdução de Sinais/genética , Via de Sinalização Wnt/genética , beta Catenina/genética
11.
Genes Dev ; 33(19-20): 1361-1366, 2019 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-31488580

RESUMO

The ubiquitin-specific protease (USP) family is the largest group of cysteine proteases. Cancer genomic analysis identified frequent amplification of USP21 (22%) in human pancreatic ductal adenocarcinoma (PDAC). USP21 overexpression correlates with human PDAC progression, and enforced expression of USP21 accelerates murine PDAC tumor growth and drives PanIN to PDAC progression in immortalized human pancreatic ductal cells. Conversely, depletion of USP21 impairs PDAC tumor growth. Mechanistically, USP21 deubiquitinates and stabilizes the TCF/LEF transcription factor TCF7, which promotes cancer cell stemness. Our work identifies and validates USP21 as a PDAC oncogene, providing a potential druggable target for this intractable disease.


Assuntos
Regulação Neoplásica da Expressão Gênica/genética , Neoplasias Pancreáticas/enzimologia , Ubiquitina Tiolesterase/genética , Ubiquitina Tiolesterase/metabolismo , Via de Sinalização Wnt/genética , Animais , Linhagem Celular Tumoral , Humanos , Camundongos , Células-Tronco Neoplásicas/patologia , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/fisiopatologia , Fator 1 de Transcrição de Linfócitos T , Ubiquitinação
12.
Nat Commun ; 10(1): 3993, 2019 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-31488837

RESUMO

Planar cell polarity (PCP) plays crucial roles in developmental processes such as gastrulation, neural tube closure and hearing. Wnt pathway mutants are often classified as PCP mutants due to similarities between their phenotypes. Here, we show that in the zebrafish lateral line, disruptions of the PCP and Wnt pathways have differential effects on hair cell orientations. While mutations in the PCP genes vangl2 and scrib cause random orientations of hair cells, mutations in wnt11f1, gpc4 and fzd7a/b induce hair cells to adopt a concentric pattern. This concentric pattern is not caused by defects in PCP but is due to misaligned support cells. The molecular basis of the support cell defect is unknown but we demonstrate that the PCP and Wnt pathways work in parallel to establish proper hair cell orientation. Consequently, hair cell orientation defects are not solely explained by defects in PCP signaling, and some hair cell phenotypes warrant re-evaluation.


Assuntos
Polaridade Celular/genética , Polaridade Celular/fisiologia , Células Ciliadas Auditivas/metabolismo , Via de Sinalização Wnt/genética , Via de Sinalização Wnt/fisiologia , Peixe-Zebra/genética , Animais , Regulação da Expressão Gênica no Desenvolvimento , Proteoglicanas de Heparan Sulfato/genética , Proteínas de Membrana/genética , Morfogênese/genética , Morfogênese/fisiologia , Mutação , Defeitos do Tubo Neural/genética , Neurulação/genética , Receptores de Superfície Celular/genética , Proteína Wnt1/genética , Proteínas de Peixe-Zebra/genética
13.
Artif Cells Nanomed Biotechnol ; 47(1): 3400-3409, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31401873

RESUMO

To grab the possible impact of lncRNA-SVUGP2 in the biology and process of non-small cell lung cancer (NSCLC). Sixty paired NSCLC tumour and the adjacent non-tumour lung tissues were collected for detection of lncRNA-SVUGP2. lncRNA-SVUGP2 expression in NSCLC cells (SK-MES-1, A549, SPC-A1, and NCI-H1975) was also detected. lncRNA-SVUGP2 was overexpressed and depressed in A549 and H1975 cells, and the effects of lncRNA-SVUGP2 dysregulation on cell biological performances including viability, colony formation, apoptosis, migration and invasion were grabbed. Furthermore, the regulatory association of lncRNA-SVUGP2 vs. EZH2 in H1975 cells, as well as the association between lncRNA-SVUGP2 and Wnt/ß-catenin pathway, was explored. lncRNA-SVUGP2 was depressed in NSCLC tissues and cells. Overexpression of lncRNA-SVUGP2 depressed proliferation, induced apoptosis, and suppressed migration and invasion of A549 and H1975 cells. In addition, lncRNA-SVUGP2 was repressed by EZH2 and was inversely correlated with EZH2 levels in H1975 cells. Repression of lncRNA-SVUGP2 potentially participated in the oncogenic function of EZH2. Besides, overexpression of lncRNA-SVUGP2 depressed the briskness of Wnt/ß-catenin signal in H1975 cells. Our data reveal that lncRNA-SVUGP2 is under-expressed in NSCLC cells and the reduced expression of lncRNA-SVUGP2 may enhance the development and process of NSCLC by interacting with EZH2 and activating Wnt/ß-catenin pathway.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/patologia , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Neoplasias Pulmonares/patologia , RNA Longo não Codificante/genética , Via de Sinalização Wnt/genética , Apoptose/genética , Carcinogênese/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Invasividade Neoplásica/genética
14.
Artif Cells Nanomed Biotechnol ; 47(1): 3278-3285, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31379206

RESUMO

Objective: The current study aimed to explore the function of miR-638 on the progression of oral squamous cell carcinoma (OSCC) and relevant molecular mechanisms. Methods: Expression profile of miR-638 in OSCC tissues and cells was detected using quantitative real-time polymerase chain reaction (qRT-PCR) method. Chi-square test was performed to estimate the relationship between miR-638 and clinical parameters of OSCC cases. Cell viability and motility abilities were estimated using MTT and transwell assays, respectively. Potential targets of miR-638 in OSCC were identified through bioinformatics analysis and luciferase reporter assay. Results: MiR-638 exhibited decreased expression in OSCC tissues and cells, compared to non-cancerous controls (P < .05 for both). Moreover, its down-regulation was closely correlated with lymph node metastasis (P = .044) and TNM stages (P = .001). Enforced miR-638 expression reduced cell proliferation, migration and invasion, while its knockdown exhibited opposite effects. Phospholipase D1 (PLD1) was confirmed as a target of miR-638 in OSCC. MiR-638 could inhibit wnt/ß-catenin pathway through targeting PLD1, thus realizing its anti-tumour action in OSCC. Conclusion: MiR-638 may be a tumour suppressor in OSCC by targeting PLD1/Wnt/ß-catenin pathway.


Assuntos
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , MicroRNAs/genética , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Fosfolipase D/genética , Via de Sinalização Wnt/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Regulação para Baixo/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/genética
15.
BMC Bioinformatics ; 20(1): 412, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31366320

RESUMO

BACKGROUND: Cartilage damage is a crucial feature involved in several pathological conditions characterized by joint disorders, such as osteoarthritis and rheumatoid arthritis. Accumulated evidences showed that Wnt/ß-catenin pathway plays a role in the pathogenesis of cartilage damage. In addition, it is experimentally documented that lncRNA (long non-coding RNA) HOTAIR plays a key role in the regulation of Wnt/ß-catenin pathway based on directly decreased WIF-1 expression. Further, it is reported that Wnt/ß-catenin pathway is a potent pathway to regulate the expression of MMP-13, which is responsible for degradation of collagen type II in articular cartilage. It is increasingly recognized that systems modeling approach provides an opportunity to understand the complex relationships and direct quantitative analysis of dynamic network in various diseases. RESULTS: A dynamic network of lncRNA HOTAIR-mediated Wnt/ß-catenin pathway regulating MMP-13 is developed to investigate the dynamic mechanism of the network involved in the pathogenesis of cartilage damage. Based on the network modeling, the potential therapeutic intervention point Axin is predicted and confirmed by the experimental validation. CONCLUSIONS: Our study provides a promising strategy for revealing potential dynamic mechanism and assessing potential targets which contribute to the prevention of the pathological conditions related to cartilage damage.


Assuntos
Cartilagem Articular/patologia , Redes Reguladoras de Genes , Terapia de Alvo Molecular , RNA Longo não Codificante/metabolismo , Via de Sinalização Wnt , Proteína Axina/farmacologia , Cartilagem Articular/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Humanos , Metaloproteinase 13 da Matriz/metabolismo , Modelos Biológicos , RNA Longo não Codificante/genética , Reprodutibilidade dos Testes , Fatores de Tempo , Via de Sinalização Wnt/efeitos dos fármacos , Via de Sinalização Wnt/genética
16.
Mol Carcinog ; 58(11): 1974-1984, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31338875

RESUMO

Wnt/ß-catenin signaling plays an important role in melanocyte biology, especially in the early stages of melanocyte transformation and melanomagenesis. ß-catenin, encoded by the gene CTNNB1, is an intracellular signal transducer of Wnt signaling and activates transcription of genes important for cell proliferation and survival. Wnt/ß-catenin signaling is frequently activated in melanoma through oncogenic mutations of ß-catenin and elevated ß-catenin levels are positively correlated with melanoma aggressiveness. Molecular mechanisms that regulate ß-catenin expression in melanoma are not fully understood. MicroRNA-214 is known to function as a tumor suppressor by targeting ß-catenin in several types of cancer cells. Here, we investigated the regulation of ß-catenin by miR-214 and its role in melanoma. We show that ß-catenin mRNA levels are negatively correlated with miR-214 in melanoma. However, overexpression of miR-214 paradoxically increased ß-catenin protein levels and promoted malignant properties of melanoma cells including resistance to mitogen-activated protein kinase inhibitors (MAPKi). RNA-seq analysis revealed that melanoma cells predominantly express a ß-catenin mRNA isoform lacking miR-214 target site. Using matched miRNA and mRNA-seq and bioinformatics analysis, we identified novel miR-214 targets, ankyrin repeat domain 6 (ANKRD6) and C-terminal binding protein 1 (CTBP1), that are involved in negative regulation of Wnt signaling. Overexpression of miR-214 or knockdown of the novel miR-214 targets, ANKRD6 or CTBP1, increased melanoma cell proliferation, migration, and decreased sensitivity to MAPKi. Our data suggest that in melanoma cells ß-catenin is not regulated by miR-214 and the functions of miR-214 in melanoma are mediated partly by regulating proteins involved in attenuation of Wnt/ß-catenin signaling.


Assuntos
Melanoma/genética , MicroRNAs/genética , beta Catenina/genética , Oxirredutases do Álcool/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Proteínas do Citoesqueleto/genética , Proteínas de Ligação a DNA/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Melanoma/patologia , RNA Mensageiro/genética , Via de Sinalização Wnt/genética
17.
Artif Cells Nanomed Biotechnol ; 47(1): 3037-3042, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31342798

RESUMO

Aim: The study aimed to investigate the role of miR-29a in the progression of oral squamous cell carcinoma (OSCC), as well as its molecular mechanism. Methods: Tissue samples were collected from 103 OSCC patients. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-29a in OSCC tissues and cell line. Cell proliferation, motility and apoptosis were detected using MTT, transwell and flow cytometry methods, respectively. Western blot was used to measure the protein expression. Results: The expression of miR-29a was decreased in OSCC tissue and cells (p < .05 for both), and its down-regulation was negatively associated with the lymph node metastasis (p = .017) and TNM stage (p = .007). Enforcing miR-29a expression in OSCC cells using mimic transfection could significantly inhibit the proliferation, migration and invasion, and promote cells apoptosis. Furthermore, miR-29a over-expression could suppress Wnt/ß-catenin pathway activity. Meanwhile, LiCl, the activator of Wnt/ß-catenin pathway, could reverse the anti-tumour action induced by miR-29a over-expression. Conclusions: MiR-29a may inhibit the malignant progression of OSCC by suppressing Wnt/ß-catenin signalling pathway.


Assuntos
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Progressão da Doença , MicroRNAs/genética , Neoplasias Bucais/genética , Neoplasias Bucais/patologia , Via de Sinalização Wnt/genética , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade
18.
Nat Commun ; 10(1): 3257, 2019 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-31332174

RESUMO

How multicellular organisms assess and control their size is a fundamental question in biology, yet the molecular and genetic mechanisms that control organ or organism size remain largely unsolved. The freshwater polyp Hydra demonstrates a high capacity to adapt its body size to different temperatures. Here we identify the molecular mechanisms controlling this phenotypic plasticity and show that temperature-induced cell number changes are controlled by Wnt- and TGF-ß signaling. Further we show that insulin-like peptide receptor (INSR) and forkhead box protein O (FoxO) are important genetic drivers of size determination controlling the same developmental regulators. Thus, environmental and genetic factors directly affect developmental mechanisms in which cell number is the strongest determinant of body size. These findings identify the basic mechanisms as to how size is regulated on an organismic level and how phenotypic plasticity is integrated into conserved developmental pathways in an evolutionary informative model organism.


Assuntos
Tamanho Corporal/fisiologia , Hydra/metabolismo , Receptor de Insulina/metabolismo , Transdução de Sinais/fisiologia , Fator de Crescimento Transformador beta/metabolismo , Via de Sinalização Wnt/fisiologia , Animais , Tamanho Corporal/genética , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Hydra/genética , Hydra/crescimento & desenvolvimento , Insulina/metabolismo , Receptor de Insulina/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Receptores de Fatores de Crescimento Transformadores beta/metabolismo , Transdução de Sinais/genética , Temperatura Ambiente , Fator de Crescimento Transformador beta/genética , Via de Sinalização Wnt/genética
19.
Cancer Sci ; 110(9): 2794-2805, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31336010

RESUMO

SALL4 is overexpressed in many cancers and is found to be involved in tumorigenesis and tumor progression. However, the function of SALL4 in cervical cancer remains unknown. Here, we showed that the expression of SALL4 was gradually increased from normal cervical tissue to high-grade squamous intraepithelial lesions and then to squamous cervical carcinoma. SALL4 was upregulated or downregulated in cervical cancer cells by stably transfecting a SALL4-expressing plasmid or a shRNA plasmid targeting SALL4, respectively. In vitro, cell growth curves and MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) assays showed that SALL4 promoted the cell proliferation of cervical cancer cells. In vivo, xenograft experiments verified that SALL4 enhanced the tumor formation of cervical cancer cells in female BALB/c Nude mice. Cell cycle analysis by fluorescence-activated cell sorting found that SALL4 accelerates cell cycle transition from the G0 /G1 phase to the S phase. TOP/FOP-Flash reporter assay revealed that SALL4 significantly upregulates the activity of Wnt/ß-catenin pathway. Western blotting showed that the expression levels of ß-catenin and important downstream genes, including c-Myc and cyclin D1, were increased by SALL4 in cervical cancer cells. Furthermore, dual-luciferase reporter and chromatin immunoprecipitation assays confirmed that SALL4 transcriptionally activated CTNNB1 by physically interacting with its promoters. Taken together, The results of this study demonstrated that SALL4 may promote cell proliferation and tumor formation of cervical cancer cells by upregulating the activity of the Wnt/ß-catenin signaling pathway by directly binding to the CTNNB1 promoter and trans-activating CTNNB1.


Assuntos
Carcinoma de Células Escamosas/genética , Transformação Celular Neoplásica/genética , Lesões Intraepiteliais Escamosas Cervicais/patologia , Fatores de Transcrição/metabolismo , Neoplasias do Colo do Útero/genética , Via de Sinalização Wnt/genética , beta Catenina/genética , Animais , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Colo do Útero/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Regiões Promotoras Genéticas/genética , RNA Interferente Pequeno/metabolismo , Lesões Intraepiteliais Escamosas Cervicais/genética , Fatores de Transcrição/genética , Regulação para Cima , Neoplasias do Colo do Útero/patologia , Ensaios Antitumorais Modelo de Xenoenxerto , beta Catenina/metabolismo
20.
Artif Cells Nanomed Biotechnol ; 47(1): 3013-3020, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31321998

RESUMO

This research planned to dig the impacts and potential principles of long noncoding RNA RP4 onH9c2 cell injury induced by hypoxia. The H9c2 cardiac muscle cells were cultured under 3% O2 concentration to induce hypoxia injury, followed by detection of RP4 expression. RP4 was then overexpressed and silenced to investigate its effects on cell injury induced by hypoxia. The potential correlation between RP4 and miR-939, between miR-939 and Bnip3, and between RP4/miR-939/Bnip3 axis and Wnt/ß-catenin pathway activation were explored. Biological processes (suppressed cell viability, migration and invasion, but enhanced cell apoptosis) were changed by hypoxia. Upregulation of RP4 enhanced hypoxia-produced damage in H9c2 cells. Additionally, miR-939 expression was opposite regulated by RP4, and miR-939 mimic abrogated the influences of pc-RP4 on enhanced hypoxia damage in H9c2 cells. Moreover, Bnip3 was targeted by miR-939 and their correlation is negative. Furthermore, upregulation of RP4 exacerbated hypoxia-produced injury in H9c2 cells by sensitizing Wnt/ß-catenin signals in H9c2 cells, which was regulated by miR-939/Bnip3 axis. Our findings reveal that RP4 is highly expressed in the hypoxia-resulted H9c2 cells. Enhanced expression of RP4 may exacerbate hypoxia injury in cardiomyocytes through regulating miR-939/Bnip3 axis-mediated briskness of Wnt/ß-catenin signals. Our study will offer a fresh theoretical basis for the treatment of ischemic myocardial injury.


Assuntos
Proteínas de Membrana/metabolismo , MicroRNAs/genética , Proteínas Mitocondriais/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , RNA Longo não Codificante/genética , Regulação para Cima , Via de Sinalização Wnt/genética , Animais , Hipóxia Celular/genética , Linhagem Celular , Ratos
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