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1.
Gene ; 721: 144097, 2019 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-31493507

RESUMO

BACKGROUND: Polo-like kinase 1 (PLK1) is a potential prognostic marker in colorectal cancer (CRC). Nevertheless, the clinicopathological and prognostic roles of PLK1 in CRC are still undefined. Therefore, we performed a meta-analysis to investigate the clinicopathological and prognostic relevance of PLK1 expression in CRC patients. METHODS: Studies published between 2003 and 2016 were selected for the meta-analysis based on an electronic literature search (PubMed, EMBASE and Chinese databases). Studies that investigated the clinicopathological and prognostic impacts of PLK1 expression in CRC patients were included for this analysis. RESULTS: Eleven studies that enrolled 1147 CRC patients were included in our meta-analysis. The effect of PLK1 level on overall survival (OS) was reported in five studies, which included 702 patients. Ten studies investigated the clinicopathological role of PLK1 expression in CRC patients. Consequently, PLK1 overexpression was associated with poorer OS in CRC patients. Furthermore, the results revealed that higher PLK1 levels were also observed in CRC tissues compared with that of normal colorectal tissues. In addition, this meta-analysis also revealed positive correlations between PLK1 upregulation and lymph node metastasis or invasion. PLK1 overexpression was significantly correlated with advanced TNM stages and higher Dukes stages. CONCLUSION: This meta-analysis strongly supports the hypothesis that PLK1 might serve as an important factor in evaluating the biological behavior and prognosis of CRC.


Assuntos
Biomarcadores Tumorais/biossíntese , Proteínas de Ciclo Celular/biossíntese , Neoplasias Colorretais , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Proto-Oncogênicas/biossíntese , Biomarcadores Tumorais/genética , Proteínas de Ciclo Celular/genética , Neoplasias Colorretais/enzimologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Intervalo Livre de Doença , Humanos , Metástase Linfática , Estadiamento de Neoplasias , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/genética , Taxa de Sobrevida
2.
Gene ; 701: 15-22, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30898709

RESUMO

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is one of the most common causes of cancer-related death. Increasing evidence suggests that cell cycle dysregulation is one of the hallmarks of cancer. In this study, by using the GEO database, we predicted the cell cycle-related protein CDK1 and BUB1 to be significantly overexpressed in PDAC tissues. Thus, this study aimed to investigate the clinical pathological significance of CDK1 and BUB1 in PDAC. METHODS: To explore the role of CDK1 and BUB1 in PDAC progression and evaluate their prognostic value, we investigated the expression patterns of CDK1 and BUB1 by using immunohistochemical staining in 99 PDAC and 71 normal pancreatic tissues with complete pathological parameters and survival data. RESULTS: CDK1 and BUB1 were significantly overexpressed in PDAC tissues. The expression of CDK1 was correlated with tumor size and histological grade, and the expression of BUB1 was correlated with the tumor size of PDAC. With regard to survival, a high expression of either CDK1 or BUB1 was correlated with a short survival of PDAC patients. Additionally, PDAC patients with a concurrent high expression of CDK1 and BUB1 showed the shortest survival. CONCLUSIONS: Our study demonstrated that CDK1 and BUB1 may play a role in PDAC progression and could be prognostic biomarkers for PDAC patients.


Assuntos
Biomarcadores Tumorais/biossíntese , Proteína Quinase CDC2/biossíntese , Carcinoma Ductal Pancreático , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/biossíntese , Neoplasias Pancreáticas , Proteínas Serina-Treonina Quinases/biossíntese , Carcinoma Ductal Pancreático/enzimologia , Carcinoma Ductal Pancreático/mortalidade , Carcinoma Ductal Pancreático/patologia , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/patologia , Valor Preditivo dos Testes , Taxa de Sobrevida
3.
Gynecol Oncol ; 153(2): 425-435, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30853360

RESUMO

OBJECTIVES: The PI3K/AKT/mTOR pathway is frequently overactivated in endometrial cancer (EC). We assessed the efficacy of ABTL0812, a novel first-in-class molecule presenting a unique mechanism of action inhibiting this pathway. METHODS: We investigated the effects of ABTL0812 on proliferation, cell death and modulation of intracellular signaling pathways in a wide panel of endometrioid and non-endometrioid cell lines, an inducible PTEN knock-out murine model, and two patient-derived xenograft murine models of EC. Then, TRIB3 expression was evaluated as potential ABTL0812 pharmacodynamic biomarker in a Phase 1b/2a clinical trial. RESULTS: ABTL0812 induced an upregulation of TRIB3 expression, resulting in the PI3K/AKT/mTOR axis inhibition and autophagy cell death induction on EC cells but not in healthy endometrial cells. ABTL0812 treatment also impaired PTEN knock-out cells to progress from hyperplasia to cancer. The therapeutic effects of ABTL0812 were demonstrated in vivo. ABTL0812 increased TRIB3 mRNA levels in whole blood samples of eight EC patients, demonstrating that TRIB3 mRNA could be used as a pharmacodynamic biomarker to monitor the ABTL0812 treatment. CONCLUSIONS: ABTL0812 may represent a novel and highly effective therapeutic agent by inducing TRIB3 expression and autophagy in EC patients, including those with poorer prognosis.


Assuntos
Antineoplásicos/farmacologia , Proteínas de Ciclo Celular/metabolismo , Neoplasias do Endométrio/tratamento farmacológico , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Repressoras/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologia , Idoso , Animais , Autofagia/efeitos dos fármacos , Proteínas de Ciclo Celular/biossíntese , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios Clínicos Fase I como Assunto , Ensaios Clínicos Fase II como Assunto , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Feminino , Humanos , Camundongos , Pessoa de Meia-Idade , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Repressoras/biossíntese , Proteínas Repressoras/genética , Regulação para Cima/efeitos dos fármacos
4.
Development ; 146(3)2019 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-30665887

RESUMO

In the Arabidopsis stomatal lineage, cells transit through several distinct precursor identities, each characterized by unique cell division behaviors. Flexibility in the duration of these precursor phases enables plants to alter leaf size and stomatal density in response to environmental conditions; however, transitions between phases must be complete and unidirectional to produce functional and correctly patterned stomata. Among direct transcriptional targets of the stomatal initiating factor SPEECHLESS, a pair of genes, SOL1 and SOL2, are required for effective transitions in the lineage. We show that these two genes, which are homologs of the LIN54 DNA-binding components of the mammalian DREAM complex, are expressed in a cell cycle-dependent manner and regulate cell fate and division properties in the self-renewing early lineage. In the terminal division of the stomatal lineage, however, these two proteins appear to act in opposition to their closest paralog, TSO1, revealing complexity in the gene family that may enable customization of cell divisions in coordination with development.


Assuntos
Arabidopsis/metabolismo , Ciclo Celular/fisiologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Estômatos de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Receptores de Superfície Celular/biossíntese , Arabidopsis/genética , Estômatos de Plantas/genética , Proteínas Serina-Treonina Quinases/genética , Receptores de Superfície Celular/genética
5.
Hum Pathol ; 83: 133-139, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30171989

RESUMO

Liver kinase B1 (LKB1) is a newly discovered tumor suppressor gene that plays a role in tumorigenesis and cancer progression. However, LKB1 expression and its precise impact on gastric cancer (GC) have not yet been elucidated. The aim of this study was to explore the significance of LKB1 expression, as well as its correlation with epithelial-mesenchymal transition (EMT) in GC. In the present study, LKB1 protein was detected in 107 GC tissue samples and adjacent paracancerous tissues by immunohistochemical staining. The relationship of LKB1 expression with clinicopathological features and its correlation with 3 EMT-related markers (E-cadherin, ß-catenin, and vimentin) in GC were analyzed. Results revealed that the expression of LKB1 was decreased in GC tissues compared with that in adjacent paracancerous tissues (P = .005). GC patients with greater invasion depth (P = .007), higher pathological TNM stage (P = .014), and lymph node metastasis (P = .026) showed lower LKB1 expression; furthermore, E-cadherin and ß-catenin expression decreased, whereas vimentin expression increased (all P < .05). Kaplan-Meier analysis indicated that the expression of LKB1, E-cadherin, ß-catenin, and vimentin, as well as differentiation, invasion, pathological TNM, and lymph node metastasis, was associated with disease-free survival (DFS) (all P < .05). Multivariate analysis also showed that LKB1 expression (hazard ratio, 0.578 [0.351-0.950]; P = .031) may be an independent factor for DFS. In conclusion, LKB1 expression was decreased in GC, and this positively correlated with EMT and a shorter DFS, suggesting that LKB1 could act as an independent factor in predicting GC progression.


Assuntos
Biomarcadores Tumorais/análise , Transição Epitelial-Mesenquimal/fisiologia , Proteínas Serina-Treonina Quinases/biossíntese , Neoplasias Gástricas/patologia , Adulto , Idoso , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteínas Serina-Treonina Quinases/análise , Estudos Retrospectivos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade
6.
J Biol Chem ; 294(3): 838-851, 2019 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-30482839

RESUMO

The pyruvate dehydrogenase complex (PDC) is a multienzyme assembly that converts pyruvate to acetyl-CoA. As pyruvate and acetyl-CoA play central roles in cellular metabolism, understanding PDC regulation is pivotal to understanding the larger metabolic network. The activity of mammalian PDC is regulated through reversible phosphorylation governed by at least four isozymes of pyruvate dehydrogenase kinase (PDK). Deciphering which kinase regulates PDC in organisms at specific times or places has been challenging. In this study, we analyzed mouse strains carrying targeted mutations of individual isozymes to explore their role in regulating PDC activity. Analysis of protein content of PDK isozymes in major metabolic tissues revealed that PDK1 and PDK2 were ubiquitously expressed, whereas PDK3 and PDK4 displayed a rather limited tissue distribution. Measurement of kinase activity showed that PDK1 is the principal isozyme regulating hepatic PDC. PDK2 was largely responsible for inactivation of PDC in tissues of muscle origin and brown adipose tissue (BAT). PDK3 was the principal kinase regulating pyruvate dehydrogenase activity in kidney and brain. In a well-fed state, the tissue levels of PDK4 protein were fairly low. In most tissues tested, PDK4 ablation had little effect on the overall rates of inactivation of PDC in kinase reaction. Taken together, these data strongly suggest that the activity of PDC is regulated by different isozymes in different tissues. Furthermore, it appears that the overall flux through PDC in a given tissue largely reflects the properties of the PDK isozyme that is principally responsible for the regulation of PDC activity in that tissue.


Assuntos
Encéfalo/enzimologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Rim/enzimologia , Proteínas Serina-Treonina Quinases/biossíntese , Complexo Piruvato Desidrogenase/biossíntese , Animais , Camundongos , Especificidade de Órgãos/fisiologia
7.
Int J Mol Med ; 43(2): 821-829, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30535427

RESUMO

One of the pathological functions of heat shock protein 22 (HSP22) is the association with inflammatory diseases and atherosclerosis. However, the effects of a high­fat diet (HFD) or oxidized low­density lipoprotein (ox­LDL) combined with atorvastatin (ATV) on HSP22 expression are entirely unknown. The present study investigated the effects of ATV on HSP22 expression in HFD­induced atherosclerotic apolipoprotein E­deficient (ApoE­/­) mice and in ox­LDL­induced human umbilical vein endothelial cells (HUVECs). Furthermore, the influence of HSP22­knockdown on the HFD- or ox­LDL­induced atherosclerotic model was also examined. It was found that HFD or ox­LDL treatment significantly increased HSP22 expression in the serum and aorta, accompanied by decreased phosphorylated (p)­endothelial nitric oxide synthase (p-eNOS) activity and activated p38 mitogen­activated protein kinase (MAPK). However, these effects were suppressed by treatment with ATV. Furthermore, HSP22-knockdown showed reduced ox­LDL­induced lesions, evidenced by increased p­eNOS activity and inactivated p38 MAPK, while suppression of cell proliferation inhibition and cell cycle arrest were also observed. Taken together, the results of this study suggest that HFD or ox­LDL increased the expression of HSP22 and p­p38 MAPK, and decreased the p­eNOS activity in vitro and in vivo, and ATV could reduce the effects by downregulating HSP22 expression.


Assuntos
Anticolesterolemiantes/farmacologia , Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Atorvastatina/farmacologia , Proteínas de Choque Térmico HSP20/biossíntese , Proteínas de Choque Térmico/biossíntese , Proteínas Musculares/biossíntese , Proteínas Serina-Treonina Quinases/biossíntese , Animais , Anticolesterolemiantes/uso terapêutico , Atorvastatina/uso terapêutico , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Dieta Hiperlipídica , Regulação para Baixo , Células Endoteliais da Veia Umbilical Humana , Humanos , Lipoproteínas LDL/metabolismo , Masculino , Camundongos , Óxido Nítrico Sintase Tipo III/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
8.
Glia ; 67(1): 68-77, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30453391

RESUMO

Multiple sclerosis (MS) is a chronic inflammatory disorder of the central nervous system (CNS), characterized by inflammation-mediated demyelination, axonal injury and neurodegeneration. The mechanisms underlying impaired neuronal function are not fully understood, but evidence is accumulating that the presence of the gliotic scar produced by reactive astrocytes play a critical role in these detrimental processes. Here, we identified astrocytic Transient Receptor Potential cation channel, subfamily M, member 7 (TRPM7), a Ca2+ -permeable nonselective cation channel, as a novel player in the formation of a gliotic scar. TRPM7 was found to be highly expressed in reactive astrocytes within well-characterized MS lesions and upregulated in primary astrocytes under chronic inflammatory conditions. TRPM7 overexpressing astrocytes impaired neuronal outgrowth in vitro by increasing the production of chondroitin sulfate proteoglycans, a key component of the gliotic scar. These findings indicate that astrocytic TRPM7 is a critical regulator of the formation of a gliotic scar and provide a novel mechanism by which reactive astrocytes affect neuronal outgrowth.


Assuntos
Astrócitos/metabolismo , Proteoglicanas de Sulfatos de Condroitina/biossíntese , Esclerose Múltipla/metabolismo , Neurônios/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Canais de Cátion TRPM/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Células Cultivadas , Proteoglicanas de Sulfatos de Condroitina/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/genética , Esclerose Múltipla/patologia , Neurônios/patologia , Proteínas Serina-Treonina Quinases/genética , Ratos , Canais de Cátion TRPM/genética
9.
Biomed Pharmacother ; 109: 679-689, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30551520

RESUMO

LATS2 is a classical tumor suppressor that affects non-small cell lung cancer proliferation and mobilization. However, its role in lung cancer cell apoptosis is unknown. The aim of our study is to explore whether LATS2 activates mitochondria-related apoptosis in lung cancer cells. In the present study, A549 non-small cell lung cancer cells were transfected with a LATS2 adenovirus to induce LATS2 overexpression. Cell apoptosis was evaluated via the MTT assay, TUNEL staining, western blotting, trypan blue staining and ELISA. Mitochondrial function was measured using an immunofluorescence assay, western blotting and ELISA. The results demonstrated that LATS2 was downregulated in A549 lung cancer cells. Overexpression of LATS2 induced A549 cell apoptosis via activating mitochondrial fission. Subsequently, we confirmed that LATS2 modulated mitochondrial fission via the JNK-Mff signaling pathway. Inhibition of the JNK pathway and/or knockdown of Mff abolished the pro-apoptotic effect of LATS2 on A549 cells. Taken together, our results identified LATS2 as a classical tumor suppressor of lung cancer via triggering mitochondrial fission and activating the JNK-Mff signaling pathway. Our results lay the foundation for detailed study of the molecular mechanisms of LATS2 overexpression and regulation of mitochondrial fission for lung cancer treatment.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas de Membrana/metabolismo , Dinâmica Mitocondrial/fisiologia , Proteínas Mitocondriais/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Supressoras de Tumor/biossíntese , Células A549 , Apoptose/fisiologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Humanos , Neoplasias Pulmonares/patologia
10.
Biomed Res Int ; 2018: 7897346, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30363964

RESUMO

Objective: To evaluate the association between upregulated differentially expressed genes (DEGs) and the outcomes of patients with hepatocellular carcinoma (HCC). Methods: Using Gene Expression Omnibus (GEO) datasets including GSE45436, GSE55092, GSE60502, GSE84402, and GSE17548, we detected upregulated DEGs in tumors. KEGG, GO, and Reactome enrichment analysis of the DEGs was conducted to clarify their function. The impact of the upregulated DEGs on patients' survival was analyzed based on TCGA profile. Results: 161 shared upregulated DEGs were identified among GSE45436, GSE55092, GSE60502, and GSE84402 profiles. Cell cycle was the shared pathway/biological process in the gene sets investigation among databases of KEGG, GO, and Reactome. After being validated in GSE17548, 13 genes including BUB1B, CCNA2, CCNB1, CCNE2, CDC20, CDC6, CDC7, CDK1, CDK4, CDKN2A, CHEK1, MAD2L1, and MCM3 in cell cycle pathway were shared in the three databases for enrichment. The expression of BUB1B, CCNB1, CDC7, CDC20, and MCM3 was upregulated in HCC tissues when compared with adjacent normal tissues in 6.67%, 7.5%, 8.06%, 5.56%, and 9.72% of HCC patients, respectively. Overexpression of BUB1B, CCNB1, CDC7, CDC20, and MCM3 in HCC tissues accounted for poorer overall survival (OS) and disease-free survival (DFS) in HCC patients (all log rank P < 0.05). BUB1B, CCNB1, CDC7, CDC20, and MCM3 were all overexpressed in HCC patients with neoplasm histologic grade G3-4 compared to those with G1-2 (all P < 0.05). BUB1B, CCNB1, and CDC20 were significantly upregulated in HCC patients with vascular invasion (all P < 0.05). Additionally, levels of BUB1B, CCNB1, CDC7, and CDC20 were significantly higher in HCC patients deceased, recurred, or progressed (all P < 0.05). Conclusion: Correlated with advanced histologic grade and/or vascular invasion, upregulation of BUB1B, CCNB1, CDC7, CDC20, and MCM3 in HCC tissues predicted worse OS and DFS in HCC patients. These genes could be novel therapeutic targets for HCC treatment.


Assuntos
Carcinoma Hepatocelular , Proteínas Cdc20/biossíntese , Proteínas de Ciclo Celular/biossíntese , Ciclina B1/biossíntese , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas , Componente 3 do Complexo de Manutenção de Minicromossomo/biossíntese , Proteínas de Neoplasias/biossíntese , Proteínas Serina-Treonina Quinases/biossíntese , Regulação para Cima , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Masculino , Taxa de Sobrevida
11.
Med Sci Monit ; 24: 7424-7430, 2018 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-30332398

RESUMO

BACKGROUND In view of the high incidence of posterior capsule opacification (PCO) and the effects of TGF-ß signaling on the epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs), our study aimed to explore the mechanism of the function of TGF-ß signaling in LECs EMT. MATERIAL AND METHODS Human lens epithelial cells (HLEC-h3) were treated with TGF-ß, ILK siRNA, ILK inhibitor, and NF-κB inhibitor to study the effects of TGF-ß, ILK, and NF-κB on cell migration and EMT. Cell migration assay was used to measure cell migration ability. Western blot was performed to detect the expression of ILK, E-cadherin, and a-SMA at the protein level. QRT-PCR was used to detect the expression of ILK at the mRNA level. RESULTS Compared with control cells, TGF-ß treatment increased the expression level of ILK HLEC-h3, promoted migration of HLEC-h3 cells, increased the expression level of E-cadherin protein, and decreased the expression level of a-SMA protein. However, treatment with ILK siRNA, ILK inhibitor, and NF-κB inhibitor reversed the effects of TGF-ß on HLEC-h3 cells. CONCLUSIONS TGF-ß-stimulated ILK regulates the migration and EMT of human LECs via NF-κB.


Assuntos
Opacificação da Cápsula/metabolismo , Cristalino/metabolismo , NF-kappa B/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Antígenos CD/metabolismo , Caderinas/metabolismo , Opacificação da Cápsula/patologia , Movimento Celular/efeitos dos fármacos , Movimento Celular/fisiologia , Células Cultivadas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Humanos , Cristalino/citologia , Cristalino/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Cápsula Posterior do Cristalino/metabolismo , Cápsula Posterior do Cristalino/patologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/biossíntese , Transdução de Sinais , Fator de Crescimento Transformador beta1/metabolismo
12.
PLoS One ; 13(9): e0203397, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30180179

RESUMO

Vaccinia-related kinase 1 (VRK1) is a pro-proliferative nuclear kinase. Mice engrafted with VRK1-depleted MDA-MB-231 breast cancer cells have been shown to develop fewer distal metastases than controls, suggesting VRK1 might play a role in cell migration, invasion, and/or colonization. In work described herein, we investigated the impact of VRK1 overexpression on human mammary epithelial cells. In 2D culture, VRK1 overexpression diminishes cell migration and invasion and impairs the migration-associated processes of cell spreading and cytoskeletal rearrangement. VRK1-overexpressing cells show reduced accumulation of the mesenchymal marker vimentin and increased accumulation of the epithelial markers E-cadherin and claudin-1. VRK1 overexpression also leads to reduced levels of the transcriptional repressors snail, slug, and twist1. Cumulatively, these data indicate that VRK1 overexpression augments the epithelial properties of both MCF10a and MDA-MB-231 cells. We further studied the impact of VRK1 on the epithelial properties of MCF10a cells in 3D matrigel culture, in which cells proliferate and form epithelial sheets that mature into hollow spherical acini. VRK1 overexpression significantly accelerates the initial stages of cell proliferation, leading to larger acini that nevertheless differentiate and mature. Our analysis of human tumor tissue microarrays (TMAs) revealed that VRK1 protein levels are higher in lymph node metastases than in patient-matched mammary tumors. Using public databases, we determined that VRK1 is among the top 10% of overexpressed transcripts in multiple subtypes of invasive breast cancer, and that high levels of VRK1 expression are correlated with decreased relapse-free survival. In sum, overexpression of VRK1, by regulating the transcription repressors snail, slug, and twist1, can promote a mesenchymal-to-epithelial transition (MET) in cell culture. VRK1-mediated MET might facilitate the colonization of distal sites by metastatic breast cancer cells, providing some insight into the frequent association of VRK1 overexpression with breast malignancies and the correlation between VRK1 overexpression and poor clinical outcome.


Assuntos
Neoplasias da Mama/enzimologia , Movimento Celular , Transição Epitelial-Mesenquimal , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , Glândulas Mamárias Humanas/enzimologia , Proteínas de Neoplasias/biossíntese , Proteínas Serina-Treonina Quinases/biossíntese , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Caderinas/biossíntese , Caderinas/genética , Linhagem Celular Tumoral , Claudina-1/biossíntese , Claudina-1/genética , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Metástase Linfática , Glândulas Mamárias Humanas/patologia , Invasividade Neoplásica , Proteínas de Neoplasias/genética , Proteínas Serina-Treonina Quinases/genética
13.
Sci Rep ; 8(1): 11265, 2018 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-30050127

RESUMO

Mutations in Vaccinia-related kinase 1 (VRK1) have emerged as a cause of severe neuronal phenotypes in human, including brain developmental defects and degeneration of spinal motor neurons, leading to Spinal Muscular Atrophy (SMA) or early onset Amyotrophic Lateral Sclerosis (ALS). Vrk1 gene-trap partial Knockout (KO) mice (Vrk1GT3/GT3), which express decreased levels of Vrk1, are sterile due to impaired gamete production. Here, we examined whether this mouse model also presents neuronal phenotypes. We found a 20-50% reduction in Vrk1 expression in neuronal tissues of the Vrk1GT3/GT3 mice, leading to mild neuronal phenotypes including significant but small reduction in brain mass and motor (rotarod) impairment. Analysis of gene expression in the Vrk1GT3/GT3 cortex predicts novel roles for VRK1 in neuronal pathways including neurotrophin signaling, axon guidance and pathways implicated in the pathogenesis of ALS. Together, our studies of the partial KO Vrk1 mice reveal that even moderately reduced levels of Vrk1 expression result in minor neurological impairment and indicate new neuronal pathways likely involving VRK1.


Assuntos
Encéfalo/patologia , Encéfalo/fisiopatologia , Técnicas de Silenciamento de Genes , Proteínas Serina-Treonina Quinases/biossíntese , Animais , Camundongos , Transtornos Motores , Tamanho do Órgão , Proteínas Serina-Treonina Quinases/genética , Teste de Desempenho do Rota-Rod
14.
Toxicol Appl Pharmacol ; 356: 90-97, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30053394

RESUMO

Cardenolides are plant-derived toxic substances. Their cytotoxicity and the underlying mechanistic signaling axes have been extensively documented, but only a few anti-viral activities of cardenolides and the associated signaling pathways have been reported. Previously, we reported that a variety of cardenolides impart anti-transmissible gastroenteritis coronavirus (TGEV) activity in swine testicular (ST) cells, through targeting of the cell membrane sodium/potassium pump, Na+/K+-ATPase. Herein, we further explore the potential signaling cascades associated with this anti-TGEV activity in ST cells. Ouabain, a representative cardenolide, was found to potently diminish TGEV titers and inhibit the TGEV-induced production of IL-6 in a dose dependent manner, with 50% inhibitory concentrations of 37 nM and 23 nM respectively. By pharmacological inhibition and gene silencing, we demonstrated that PI3K_PDK1_RSK2 signaling was induced in TGEV-infected ST cells, and ouabain imparted a degree of anti-TGEV activity via further augmentation of this existing PI3K_PDK1 axis signaling, in a manner dependent upon its association with the Na+/K+-ATPase. Finally, inhibition of PI3K by LY294002 or PDK1 by BX795 antagonized the anti-viral activity of ouabain and restored the TGEV virus titer and yields. This finding is the first report of a PI3K_PDK1 signaling axis further induced by ouabain and implicated in the suppression of TGEV activity and replication; greatly illuminates the underlying mechanism of cardenolide toxicity; and is expected to result in one or more anti-viral applications for the cardenolides in the future.


Assuntos
Antivirais/farmacologia , Coronavirus/efeitos dos fármacos , Ouabaína/farmacologia , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/biossíntese , Transdução de Sinais/efeitos dos fármacos , ATPase Trocadora de Sódio-Potássio/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Animais , Linhagem Celular , Cromonas/farmacologia , Replicação do DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Inativação Gênica , Interleucina-6/antagonistas & inibidores , Interleucina-6/biossíntese , Camundongos , Morfolinas/farmacologia , Proteínas Serina-Treonina Quinases/genética , Pirimidinas/farmacologia , Tiofenos/farmacologia
15.
Nature ; 559(7713): 211-216, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29973724

RESUMO

Liquid-liquid phase separation has been shown to underlie the formation and disassembly of membraneless organelles in cells, but the cellular mechanisms that control this phenomenon are poorly understood. A prominent example of regulated and reversible segregation of liquid phases may occur during mitosis, when membraneless organelles disappear upon nuclear-envelope breakdown and reappear as mitosis is completed. Here we show that the dual-specificity kinase DYRK3 acts as a central dissolvase of several types of membraneless organelle during mitosis. DYRK3 kinase activity is essential to prevent the unmixing of the mitotic cytoplasm into aberrant liquid-like hybrid organelles and the over-nucleation of spindle bodies. Our work supports a mechanism in which the dilution of phase-separating proteins during nuclear-envelope breakdown and the DYRK3-dependent degree of their solubility combine to allow cells to dissolve and condense several membraneless organelles during mitosis.


Assuntos
Mitose , Organelas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Ciclossomo-Complexo Promotor de Anáfase/metabolismo , Citoplasma/metabolismo , Grânulos Citoplasmáticos/metabolismo , Células HEK293 , Células HeLa , Humanos , Membrana Nuclear/metabolismo , Proteína I de Ligação a Poli(A)/metabolismo , Ligação Proteica , Transporte Proteico , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/biossíntese , Solubilidade , Fuso Acromático/metabolismo , Estresse Fisiológico
16.
Br J Cancer ; 118(12): 1617-1627, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29867225

RESUMO

BACKGROUND: Constitutively activated nuclear factor kappa B (NF-κB) signalling plays vital roles in bladder urothelial carcinoma (BC) progression. We investigate the effect of receptor-interacting protein kinase 4 (RIPK4) on NF-κB activation and BC progression. METHODS: The expression of RIPK4 was examined in 25 cryopreserved paired bladder samples and 112 paraffin BC specimens. In vivo and in vitro assays were performed to validate effect of RIPK4 on NF-κB pathway-mediated BC progression. RESULTS: High expression of RIPK4 was observed in BC tissues and was an independent predictor for poor overall survival. Up or downregulating the expression of RIPK4 enhanced or inhibited, respectively, the migration and invasion of BC cells in vitro and in vivo. Mechanistically, RIPK4 promoted K63-linked polyubiquitination of tumour necrosis factor receptor-associated factor 2 (TRAF2), receptor-interacting protein (RIP) and NF-κB essential modulator (NEMO). RIPK4 also promoted nuclear localisation of NF-κB-p65, and maintained activation of NF-κB substantially, leading to upregulation of VEGF-A, ultimately promoting BC cell aggressiveness. CONCLUSIONS: Our data highlighted the molecular aetiology and clinical significance of RIPK4 in BC: upregulation of RIPK4 contributes to NF-κB activation, and upregulates VEGF-A, and BC progression. Targeting RIPK4 might represent a new therapeutic strategy to improve survival for patients with BC.


Assuntos
NF-kappa B/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Neoplasias da Bexiga Urinária/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Transição Epitelial-Mesenquimal , Humanos , Estimativa de Kaplan-Meier , Invasividade Neoplásica , Metástase Neoplásica , Inclusão em Parafina , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/genética , Transdução de Sinais , Regulação para Cima , Neoplasias da Bexiga Urinária/enzimologia , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/patologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/genética
17.
Eur Rev Med Pharmacol Sci ; 22(12): 3713-3718, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29949144

RESUMO

OBJECTIVE: CircRNAs have been recently identified as important regulators in tumors biological functions. However, the clinical significance of circHIPK3 in epithelial ovarian cancer (EOC) remains unknown. PATIENTS AND METHODS: The expression of circHIPK3 in EOC tumor tissues and adjacent noncancerous tissues was analyzed by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). The association between circHIPK3 expression and clinicopathological factors was analyzed by using Chi-square test. Kaplan-Meier method and log-rank test were used to analyze the association of circHIPK3 expression with disease-free survival (DFS) and overall survival (OS) time of EOC patients. Univariate and multivariate Cox analysis was also performed. RESULTS: We found that circHIPK3 was higher expressed in EOC tissues and cells compared to adjacent normal tissue and ovarian epithelium cell line, respectively. Higher circHIPK3 expression associated with lymph node invasion, FIGO stage, and worse DFS and OS of patients. Moreover, multivariate Cox analysis showed that higher circHIPK3 was an independent predictor of DFS and OS in EOC patients. CONCLUSIONS: Thus, circHIPK3 may be a novel biomarker for predicting EOC prognosis.


Assuntos
Carcinoma Epitelial do Ovário/diagnóstico , Carcinoma Epitelial do Ovário/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/biossíntese , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , RNA/biossíntese , Adulto , Carcinoma Epitelial do Ovário/genética , Linhagem Celular Tumoral , Intervalo Livre de Doença , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Pessoa de Meia-Idade , Neoplasias Ovarianas/genética , Prognóstico , Proteínas Serina-Treonina Quinases/genética , RNA/genética , Regulação para Cima/fisiologia
18.
Mol Med Rep ; 18(1): 105-112, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29749476

RESUMO

The present study aimed to explore the effects and underlying mechanisms of microRNA (miR)­23a on pancreatic cancer (PC) cells progression. Reverse transcription­quantitative polymerase chain reaction and western blot analysis were used to detect the mRNA and protein miR­23a and PLK­1 level. Cell viability, cell cycle, migration and invasion assasy, and in vivo tumorigenicity assay were used to investigate the effects of miR­204. Further luciferase reporter assay was used to explore the mechanisms contributing to miR­204 effects. It was observed that miR­23a expression was upregulated and negatively associated with polo­like kinase­1 (PLK­1) expression in human PC tissues. PLK­1 was a direct target of miR­23a in PC cells. Functional analysis demonstrated that miR­23a overexpression suppressed cell proliferation, inhibited cell migration and invasion and promoted cell apoptosis in vitro. When PC cells were transfected with has­miR­23a PLK­1 was downregulated and its downstream molecules were deregulated, including decreased expression of B­cell lymphoma­2, cyclin B1 and vimentin, and increased expression of Bax and E­cadherin. The inhibitory effect of miR­23a on PC cell progression was observed in vivo tumor xenografts. The results of the study suggest that miR­23a inhibits PC cell progression by directly targeting PLK­1­associated signaling and promoting miR­23a expression may be a potential method for treating patients with PC.


Assuntos
Proteínas de Ciclo Celular/biossíntese , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , MicroRNAs/metabolismo , Neoplasias Pancreáticas/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Proto-Oncogênicas/biossíntese , RNA Neoplásico/metabolismo , Animais , Apoptose , Proteínas de Ciclo Celular/genética , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/genética , RNA Neoplásico/genética
19.
Biomacromolecules ; 19(6): 2320-2329, 2018 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-29767505

RESUMO

Antibody fragment (Fab')-installed polyion complex (PIC) micelles were constructed to improve targetability of small interfering RNA (siRNA) delivery to pancreatic cancer cells. To this end, we synthesized a block copolymer of azide-functionalized poly(ethylene glycol) and poly(l-lysine) and prepared PIC micelles with siRNA. Then, a dibenzylcyclooctyne (DBCO)-modified antihuman tissue factor (TF) Fab' was conjugated to azido groups on the micellar surface. A fluorescence correlation spectroscopic analysis revealed that 1, 2, or 3 molecule(s) of Fab'(s) were installed onto one micellar nanoparticle according to the feeding ratio of Fab' (or DBCO) to micelle (or azide). The resulting micelles exhibited ∼40 nm in hydrodynamic diameter, similar to that of the parent micelles before Fab' conjugation. Flow cytometric analysis showed that three molecules of Fab'-installed PIC micelles (3(Fab')-micelles) had the highest binding affinity to cultured pancreatic cancer BxPC3 cells, which are known to overexpress TF on their surface. The 3(Fab')-micelles also exhibited the most efficient gene silencing activity against polo-like kinase 1 mRNA in the cultured cancer cells. Furthermore, the 3(Fab')-micelles exhibited high penetrability and the highest cellular internalization amounts in BxPC3 spheroids compared with one or two molecule(s) of Fab'-installed PIC micelles. These results demonstrate the potential of anti-TF Fab'-installed PIC micelles for active targeting of stroma-rich pancreatic tumors.


Assuntos
Anticorpos Antineoplásicos , Proteínas de Ciclo Celular/antagonistas & inibidores , Sistemas de Liberação de Medicamentos , Inativação Gênica , Fragmentos Fab das Imunoglobulinas , Micelas , Neoplasias Pancreáticas/tratamento farmacológico , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , RNA Interferente Pequeno , Tromboplastina/antagonistas & inibidores , Anticorpos Antineoplásicos/química , Anticorpos Antineoplásicos/farmacologia , Proteínas de Ciclo Celular/biossíntese , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Humanos , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/farmacologia , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Polietilenoglicóis/química , Polietilenoglicóis/farmacologia , Polilisina/química , Polilisina/farmacologia , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas/genética , RNA Interferente Pequeno/química , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Tromboplastina/metabolismo
20.
J Enzyme Inhib Med Chem ; 33(1): 920-935, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29768059

RESUMO

Pentabromopseudilin (PBrP) is a marine antibiotic isolated from the marine bacteria Pseudomonas bromoutilis and Alteromonas luteoviolaceus. PBrP exhibits antimicrobial, anti-tumour, and phytotoxic activities. In mammalian cells, PBrP is known to act as a reversible and allosteric inhibitor of myosin Va (MyoVa). In this study, we report that PBrP is a potent inhibitor of transforming growth factor-ß (TGF-ß) activity. PBrP inhibits TGF-ß-stimulated Smad2/3 phosphorylation, plasminogen activator inhibitor-1 (PAI-1) protein production and blocks TGF-ß-induced epithelial-mesenchymal transition in epithelial cells. PBrP inhibits TGF-ß signalling by reducing the cell-surface expression of type II TGF-ß receptor (TßRII) and promotes receptor degradation. Gene silencing approaches suggest that MyoVa plays a crucial role in PBrP-induced TßRII turnover and the subsequent reduction of TGF-ß signalling. Because, TGF-ß signalling is crucial in the regulation of diverse pathophysiological processes such as tissue fibrosis and cancer development, PBrP should be further explored for its therapeutic role in treating fibrotic diseases and cancer.


Assuntos
Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Miosina Tipo V/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Pirróis/farmacologia , Receptores de Fatores de Crescimento Transformadores beta/antagonistas & inibidores , Fator de Crescimento Transformador beta/antagonistas & inibidores , Alteromonas/química , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Células HEK293 , Células Hep G2 , Humanos , Vison , Estrutura Molecular , Miosina Tipo V/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Pseudomonas/química , Pirróis/química , Pirróis/isolamento & purificação , Receptor do Fator de Crescimento Transformador beta Tipo II , Receptores de Fatores de Crescimento Transformadores beta/biossíntese , Relação Estrutura-Atividade , Fator de Crescimento Transformador beta/metabolismo
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