RESUMEN
Gastric cancer (GC) is one of the most common malignancies, leading to millions of deaths each year. Here, we investigated the molecular mechanisms of GC, with a focus on circXRCC5/miR-655-3p/RREB1/UBA2 axis. circXRCC5 was identified in 62 paired cancer specimens and adjacent normal tissues by genome-wide bioinformatics analysis and verified by qRT-PCR and Sanger sequencing. Knockdown or exogenous expression of circXRCC5 was performed to validate the functional significance of circXRCC5 using both in vitro and in vivo assays, including CCK-8, colony formation, EdU incorporation, transwell system, as well as animal experiments. RNA immunoprecipitation, biotinylated RNA pull-down, ChIP, and dual-luciferase assays were employed to validate the regulatory network of circXRCC5/miR-655-3p/RREB1/UBA2. Frequently elevated circXRCC5 in GC tissues and cell lines was associated with poor prognosis of GC patients. Functionally, circXRCC5 overexpression facilitated GC cell proliferation, migration, and invasion, as well as promoted tumor growth and metastasis in vivo. Mechanistically, circXRCC5 served as a sponge of miR-655-3p to induce upregulation of RREB1. RREB1 was identified as a transcriptional activator of UBA2, thus contributing to GC tumorigenesis. Moreover, RNA binding protein (RBP) HNRNPC was proved to interact with circXRCC5 to promote circXRCC5 biogenesis. Collectively, circXRCC5 facilitates GC progression through the HNRNPC/circXRCC5/miR-655-3p/RREB1/UBA2 axis, which might bring novel therapeutic strategies for GC treatment.
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MicroARNs , Neoplasias Gástricas , Animales , Neoplasias Gástricas/patología , MicroARNs/genética , MicroARNs/metabolismo , Regulación Neoplásica de la Expresión Génica , Proliferación Celular/genética , Retroalimentación , Línea Celular TumoralRESUMEN
A series of structurally modified curcumol derivatives at C-8 position were designed and synthesized, whose structures were confirmed by 1H NMR,13C NMR, and HRMS analysis. The tested compounds were evaluated for in vitro antitumor activity against colorectal cancer cell lines SW620, HCT116, and CaCo2. Many of the tested candidates exhibited higher inhibition efficiency than curcumol. Among them, compound 3 l shows the best inhibitory effect on the viability of SW620 with IC50 value of 19.90 ± 0.64 µM. The structure-activity relationships of these derivatives were discussed, which showed that the introduction of amino or aryl groups tended to increase the anti-cancer activity. In addition, compound 3 l may inhibit cancer cell proliferation through triggering cell apoptosis.
RESUMEN
A new series of C-14 curcumol derivatives as potent anticancer agents were designed and synthesized by click reaction, whose structures were confirmed by 1H NMR,13C NMR, and HRMS analysis. All the synthesized compounds were evaluated for in vitro antitumor activity against colorectal cancer cell lines SW620 and HCT116. Most of them exhibited higher inhibitory activity than curcumol. Especially, compound 3j shows good inhibitory activity against SW620 with IC50 value of 8.10 ± 0.13 µM. The structure-activity relationships (SARs) of these derivatives were discussed. In addition, flow cytometry revealed that compound 3j induced SW620 cells apoptosis by facilitating apoptosis-related proteins expressions. Our findings suggested that fluorine functional group on phenyl ring tended to increase the anticancer activity.
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Antineoplásicos , Sesquiterpenos , Antineoplásicos/química , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Estructura Molecular , Sesquiterpenos/farmacología , Relación Estructura-ActividadRESUMEN
Small ubiquitinlike modifier proteins are involved in tumorigenesis; however, the potential effects and functions of the family member ubiquitinlike modifieractivating enzyme 2 (UBA2) on colorectal cancer are not clear. The present study aimed to examine the effects of UBA2 on the proliferation of colorectal cancer cells in vitro and in vivo. The mRNA and protein expression levels of UBA2 in patients with colorectal cancer were measured by reverse transcriptionquantitative polymerase chain reaction and immunohistochemistry, respectively. UBA2 expression levels in colorectal cancer tissues were significantly increased compared with the paracancerous normal tissues. The expression of UBA2 was also associated with higher stage colorectal cancer and poor prognosis. MTT and colony formation assays were used to examine proliferation in colorectal cancer cell lines. Flow cytometry was performed to examine the effects of UBA2 on the cell cycle and apoptosis of colorectal cancer cell lines and protein expression levels were examined by western blotting. Athymic nude mice were used to examine the ability of transfected colorectal cancer cells to form tumors in vivo. Downregulation of UBA2 inhibited the proliferation of colorectal cancer cell lines in vitro and in vivo through the regulation of cell cycle associated protein expression and apoptosis. Furthermore, downregulation of UBA2 decreased the expression levels of cyclin B1, Bcell lymphoma-2, phosphorylated protein kinase B and E3 ubiquitinprotein ligase MDM2 in colorectal cancer cells, whereas the expression levels of p21 and p27 were increased. UBA2 was demonstrated to serve an essential role in the proliferation of colorectal cancer and may be used as a potential biomarker to predict prognosis and as a therapeutic target in colorectal cancer.
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Neoplasias Colorrectales/genética , Enzimas Activadoras de Ubiquitina/genética , Adulto , Anciano , Animales , Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/mortalidad , Neoplasias Colorrectales/patología , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Humanos , Masculino , Ratones , Persona de Mediana Edad , Clasificación del Tumor , Metástasis de la Neoplasia , Estadificación de Neoplasias , Pronóstico , Enzimas Activadoras de Ubiquitina/metabolismoRESUMEN
AIM: To investigate the function and mechanism of ubiquitin-like modifier activating enzyme 2 (Uba2) in progression of gastric cancer (GC) cells. METHODS: Uba2 level in patients with GC was analyzed by Western blotting and immunohistochemistry. MTT and colony formation assays were performed to examine cell proliferation. Flow cytometry was used for cell cycle analysis. Wound healing and Transwell assays were conducted to examine the effects of Uba2 on migration and invasion. Expression levels of cell cycle-related proteins, epithelial-mesenchymal transition (EMT) biomarkers, and involvement of the Wnt/ß-catenin pathway was assessed by Western blotting. Activation of the Wnt/ß-catenin pathway was confirmed by luciferase assay. RESULTS: Uba2 expression was higher in GC than in normal tissues. Increased Uba2 expression was correlated with tissue differentiation, Lauren's classification, vascular invasion, and TNM stage, as determined by the analysis of 100 GC cases (P < 0.05). Knock-down of Uba2 inhibited GC cell proliferation, induced cell cycle arrest, and altered expression of cyclin D1, P21, P27, and Bcl-2, while up-regulation of Uba2 showed the opposite effects. The wound healing and Transwell assays showed that Uba2 promoted GC cell migration and invasion. Western blotting revealed alterations in EMT biomarkers, suggesting the role of Uba2 in EMT. Furthermore, the luciferase reporter assay indicated the involvement of the Wnt/ß-catenin signaling pathway as a possible modulator of Uba2 oncogenic functions. CONCLUSION: Uba2 plays a vital role in GC cell migration and invasion, possibly by regulating the Wnt/ß-catenin signaling pathway and EMT.
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Movimiento Celular , Neoplasias Gástricas/patología , Enzimas Activadoras de Ubiquitina/metabolismo , Vía de Señalización Wnt , Línea Celular Tumoral , Proliferación Celular/genética , Progresión de la Enfermedad , Transición Epitelial-Mesenquimal , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Masculino , Persona de Mediana Edad , Invasividad Neoplásica/patología , Estómago/patología , Regulación hacia ArribaRESUMEN
BACKGROUND: Matrix metalloproteinase 11 (MMP11) has been shown to play a key role in human tumor progression and indicates poor clinical outcome in cancer patients. OBJECTIVE: The current study aimed to evaluate the relationship between serum levels of MMP-11 and prognosis in colon cancer patients. METHODS: Serum levels of MMP-11 were determined in 92 colon cancer patients and 92 healthy individuals using an enzyme-linked immunosorbent assay (ELISA). Associations between serum MMP-11 levels and clinicopathological characteristics of the patients and their outcomes were investigated. Survival analyses were performed to measure the 5-year overall survival (OS) and disease-free survival (DFS). RESULTS: Serum MMP-11 levels were substantially higher in colon cancer patients than in healthy controls. Moreover, serum MMP-11 levels were significantly higher in patients with advanced T status, lymph node metastasis, distant metastasis, and a higher TNM stage. Elevated serum levels of MMP-11 were identified as an independent prognostic factor for 5-year mortality and adverse events associated with colon cancer. Multivariate Cox regression analysis identified the serum MMP-11 level as an independent predictor of OS and DFS. CONCLUSION: Our study established that high serum levels of MMP-11 are associated with poor clinical outcome and may serve as a prognostic biomarker in colon cancer patients.
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Biomarcadores de Tumor , Neoplasias del Colon/sangre , Neoplasias del Colon/mortalidad , Metaloproteinasa 11 de la Matriz/sangre , Adulto , Anciano , Estudios de Casos y Controles , Neoplasias del Colon/diagnóstico , Neoplasias del Colon/tratamiento farmacológico , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Metástasis de la Neoplasia , Estadificación de Neoplasias , Pronóstico , Curva ROCRESUMEN
Inflammatory responses play critical roles in carbon monoxide (CO) poisoning-induced cerebral injury. The present study investigated whether erythropoietin (EPO) modulates the toll-like receptor 4 (TLR4) and nuclear factor-kappa B (NF-κB) inflammatory signaling pathways in brain injury after acute CO poisoning. EPO (2500 and 5000 U/kg) was injected subcutaneously twice a day after acute CO poisoning for 2 days. At 48 h after treatment, the expression levels of TLR4 and NF-κB as well as the levels of inflammatory cytokines in the hippocampal tissues were measured. Our results showed that CO poisoning induced a significant upregulation of TLR4, NF-κB, and inflammatory cytokines in the injured rat hippocampal tissues. Treatment with EPO remarkably suppressed the gene and protein expression levels of TLR4 and NF-κB, as well as the concentrations of TNF-α, IL-1ß, and IL-6 in the hippocampal tissues. EPO treatment ameliorated CO poisoning-induced histological edema and neuronal necrosis. These results suggested that EPO protected against CO poisoning-induced brain damage by inhibiting the TLR4-NF-κB inflammatory signaling pathway.
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Lesiones Encefálicas/patología , Intoxicación por Monóxido de Carbono/patología , Monóxido de Carbono/toxicidad , Edema/prevención & control , Eritropoyetina/farmacología , FN-kappa B/antagonistas & inhibidores , Necrosis/prevención & control , Receptor Toll-Like 4/antagonistas & inhibidores , Animales , Lesiones Encefálicas/inducido químicamente , Hipocampo/metabolismo , Inflamación/patología , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismoAsunto(s)
Errores Diagnósticos , Gastroparesia/diagnóstico , Hemangioma Cavernoso/complicaciones , Neoplasias del Yeyuno/complicaciones , Vómitos/diagnóstico , Diagnóstico Diferencial , Enteroscopía de Doble Balón/métodos , Femenino , Hemangioma Cavernoso/diagnóstico , Hemangioma Cavernoso/cirugía , Humanos , Neoplasias del Yeyuno/diagnóstico , Neoplasias del Yeyuno/cirugía , Laparotomía , Persona de Mediana Edad , Tomografía Computarizada por Rayos X , Vómitos/etiología , Vómitos/cirugíaRESUMEN
The jumping spider genus Epeus Peckham & Peckham presently includes 15 species, mainly from South and Southeast Asia (World Spider Catalog 2015). Species of this genus have the cymbium of male palp flattened and elongated, with a basal apophysis retrolaterally, pointing postero-ventrally; tegulum with a tongue-like process; filiform embolus; and epigyne with long copulatory ducts with several loops.
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Arañas/clasificación , Distribución Animal , Estructuras Animales/anatomía & histología , Estructuras Animales/crecimiento & desarrollo , Animales , Tamaño Corporal , China , Femenino , Masculino , Tamaño de los Órganos , Arañas/anatomía & histología , Arañas/crecimiento & desarrolloRESUMEN
BACKGROUND/AIMS: Alcohol abuse is becoming an increasingly severe problem among the Han, Mongol and Chaoxian nationalities in the northeast of China. The study aimed to investigate the relationship between alcoholic liver disease (ALD) and the genetic polymorphism of two enzymes, cytochrome P450IIE1 (CYPIIE1) and glutathione S-transferase P1 (GSTP1) in patients of three nationalities. METHODOLOGY: Peripheral blood was collected from 353 Chinese patients with ALD, 300 alcohol-dependent patients without liver disease (alcoholic) and 360 healthy controls. Each group included patients from the Han, Mongol and Chaoxian nationalities. PCR-restriction fragment length polymorphism (PCR-RFLP) was used in this research. RESULTS: Regardless of nationality patients who carried the rare CYPIIE1 C2 and GSTP1 Val allele were at higher risk of ALD. The frequency of C2 and Val in patients with ALD was 50.00% and 26.98% in the Han, 31.36% and 22.87% in the Mongol and 45.87% and 22.02% in the Chaoxian, respectively. No significant differences were seen in the frequency of either the C2 or Val alleles in ALD, among the three nationalities. In each nationality, the frequency of both the C2 and Val alleles was significantly higher in ALD compared to alcoholic and healthy controls. CONCLUSIONS: In this group of Chinese patients, we found that regardless of nationality, patients with ALD tended to carry the C2 allele and the Val allele. This may indicate a causal relationship between these polymorphic alleles that lead to the development of ALD.
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Citocromo P-450 CYP2E1/genética , Gutatión-S-Transferasa pi/genética , Hepatopatías Alcohólicas/genética , Polimorfismo Genético , Adulto , Pueblo Asiatico/genética , China/epidemiología , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Hepatopatías Alcohólicas/enzimología , Hepatopatías Alcohólicas/etnología , Masculino , Persona de Mediana Edad , Fenotipo , Reacción en Cadena de la Polimerasa , Medición de Riesgo , Factores de RiesgoRESUMEN
BACKGROUND/AIMS: Abnormalities in cell cycle regulation are reported to be strongly associated with tumorigenesis and progression of tumors. Wnt/ß-catenin signaling pathway and cell cycle play key roles during the genesis and development of hepatocellular carcinoma (HCC). Current studies indicated that expressions of cyclin A, E and D1 were affected after silencing of ß-catenin gene in HCC, but it is unclear if other cyclins are affected. METHODOLOGY: To determine the relation, small interference RNA (siRNA) against ß-catenin was transfected into HCC cell lines HepG2 and SMMC-7721, and cell cycle and cyclin B1 and cyclin C protein expression were detected. RESULTS: Cell cycle was arrested in G0/G1 at 72h after transfection and the cell cycle began to transfer from G0/G1 to G2/M through S and had a trend to revert at 96h. In addition, ß-catenin protein expression was decreased at both 72 and 96h, although the level was slightly higher at 96h than that at 72h. However, cyclin B1 expression decreased at 72h and increased at 96h, cyclin C expression increased at 72h and decreased at 96h. CONCLUSIONS: These findings suggest that silencing ß-catenin gene may induce the changes of cell cycle and cyclin B1 and cyclin C protein expression. Wnt/ß-catenin signaling pathway probably takes part in the genesis and development of HCC through regulating cell cycle and the expression of cyclin B1 and cyclin C.
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Carcinoma Hepatocelular/metabolismo , Ciclo Celular , Ciclina B1/metabolismo , Ciclina C/metabolismo , Neoplasias Hepáticas/metabolismo , Interferencia de ARN , beta Catenina/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patología , Ciclina B1/genética , Ciclina C/genética , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , Factores de Tiempo , Transfección , Vía de Señalización Wnt , beta Catenina/genéticaRESUMEN
To investigate the relationship and significance of Wnt/b-catenin signaling pathway with caspase-3, XIAP, HSP27and Grp-78. The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against b-catenin. After 72 and 96 h, protein was extracted and the protein expressions of b-catenin, caspase-3, XIAP, Grp-78 and HSP27 were detected by Western blot. b-catenin protein expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (F = 160.72, P is less than to 0.01). Interestingly, Caspase-3 protein expression was decreased at 72 h and increased to normal at 96 h (F = 136.10, P is less than to 0.01), while p-caspase-3 protein expression increased at 72 h and decreased to normal at 96 h (F = 98.65, P is less than to 0.01). XIAP protein expression decreased at 72 h (F = 37.29, P is less than to 0.01) and increased at 96 h. Grp-78 protein expression increased at 72 h and decreased to normal at 96 h ( F = 58.72, P is less than to 0.01). HSP27 protein expression showed no change following transfection ( F = 1.91, P is more than to 0.05). Wnt/b-catenin signaling pathway is related to the protein expressions of caspase-3, XIAP and Grp-78, but not related to HSP27 protein expression in HCC. Wnt/b-catenin signaling pathway may participate in the regulation of HCC apoptosis, proliferation and differentiation through affecting these factors.
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Carcinoma Hepatocelular , Caspasa 3 , Cateninas , Humanos , Neoplasias Hepáticas , Vía de Señalización Wnt , beta Catenina/metabolismoRESUMEN
BACKGROUND/AIMS: To study the correlation and significance of beta-catenin, STAT3 and GSK-3beta signaling pathway in hepatocellular carcinoma (HCC). METHODOLOGY: The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against 8-catenin or STAT3. After 72 and 96h, protein was extracted and the protein expression of beta-catenin, STAT3, and GSK-3beta was detected by Western blot analysis. RESULTS: After siRNA directed against beta-catenin was transfected into HepG2 cells, beta-catenin protein expression was decreased at 72 and 96h, GSK-3beta and p-GSK-3beta protein expression increased gradually at 72 and 96h, and STAT3 protein expression showed no change following transfection. After siRNA directed against STAT3 was transfected into HepG2 cells, STAT3 protein expression was decreased at 72 and 96h and beta-catenin, GSK-3beta and p-GSK-3beta protein expression all increased at 72h and decreased at 96 h after transfection. CONCLUSION: In HCC, the beta-catenin signaling pathway may regulate GSK-3beta protein expression and the STAT3 signaling pathway may regulate beta-catenin and GSK-3beta protein expression, thereby playing key roles during HCC genesis and development.
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Carcinoma Hepatocelular/metabolismo , Glucógeno Sintasa Quinasa 3/análisis , Neoplasias Hepáticas/metabolismo , Factor de Transcripción STAT3/fisiología , Transducción de Señal/fisiología , beta Catenina/fisiología , Carcinoma Hepatocelular/etiología , Glucógeno Sintasa Quinasa 3/fisiología , Glucógeno Sintasa Quinasa 3 beta , Células Hep G2 , Humanos , Neoplasias Hepáticas/etiología , Interferencia de ARN , Factor de Transcripción STAT3/análisis , Factor de Transcripción STAT3/genética , beta Catenina/análisis , beta Catenina/genéticaRESUMEN
BACKGROUND/PURPOSE: Bone marrow mononuclear cell (BMMC) transplantation has been shown to facilitate tissue and organ regeneration and repair. BMMC transplantation may be a potential therapy for acute liver failure, and its effect might be further improved. Hepatocyte growth factor (HGF) plays an important role in liver cell development, and may ameliorate hepatic fibrosis or cirrhosis in animal models. We therefore explored a potential synergistic effect of the co-application of HGF and BMMCs in liver regeneration following carbon tetrachloride (CCl(4))-induced acute hepatic injury. METHODS: We established a murine acute liver failure model induced by CCl(4) administration, and studied the effect of BMMC transplantation in combination with HGF. We used 4 groups of animals, one group was transfused with PKH26-labeled BMMCs (5 × 10(6)) and HGF [50 ng/(kg days) × 7 days] (BMMCs + HGF group), one group received BMMCs only, one group received HGF only, and one group received saline solution (0.9% NaCl) alone. The effects were examined by biochemical measurements of liver enzymes and quantitative image analysis for PKH26 labeling, and by determining proliferating cell nuclear antigen (PCNA) and albumin expression 4 weeks after the BMMC transplantation. RESULTS: PKH26-labeled BMMCs were detected in transplanted mouse livers, most of which expressed PCNA. PCNA and albumin expressions were increased significantly in the BMMCs + HGF group compared with the expressions of these parameters in the other 3 groups. Liver function, reflected by serum aminotransferase activity, was also improved in the BMMCs + HGF group relative to that in the other groups. CONCLUSIONS: Data from the present study appear to suggest that BMMC transplantation combined with HGF administration exhibits a synergistic beneficial effect on improving both functional and histological liver recovery in a mouse model of acute liver failure.
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Trasplante de Médula Ósea/métodos , Factor de Crecimiento de Hepatocito/metabolismo , Leucocitos Mononucleares/citología , Fallo Hepático Agudo/metabolismo , Regeneración Hepática/fisiología , Animales , Modelos Animales de Enfermedad , Femenino , Citometría de Flujo , Inmunohistoquímica , Fallo Hepático Agudo/patología , Fallo Hepático Agudo/terapia , Masculino , Ratones , Ratones Endogámicos BALB C , Resultado del TratamientoRESUMEN
OBJECTIVE: To investigate the role and significance of Wnt/beta-catenin signaling pathway regulating GSK-3beta, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC). METHODS: The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against beta-catenin. Proteins were extracted and the expressions of beta-catenin, GSK-3beta, p-GSK-3beta, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection. RESULTS: beta-catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P < 0.05). Interestingly, GSK-3beta and p-GSK-3beta expressions increased gradually at 72 and 96 h (tGSK-3beta= 4.98, tp-GSK-3beta= 29.83, P < 0.05) respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P > 0.05). Smad3 expression was increased at 72 h (t = 10.67, P < 0.05) and decreased to normal at 96 h (t = 1.26, P < 0.05), while TERT expression decreased at 72 h (t = 4.18, P is less than 0.05) and increased to normal at 96 h (t = 1.26, P > 0.05). CONCLUSIONS: Wnt/beta-catenin signaling pathway is related to the expressions of GSK-3beta, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/beta-catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.
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Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Transducción de Señal , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Carcinoma Hepatocelular/patología , Células Hep G2 , Humanos , Neoplasias Hepáticas/patología , ARN Interferente PequeñoRESUMEN
The molecular mechanism responsible for hepatocellular carcinoma (HCC) development remains to be defined although a number of gene pathways have been shown to play an active role, such as Wnt/beta-catenin signaling. In this study, beta-catenin small interfering RNA (siRNA) was designed, synthesized, and transfected into HCC HepG2 cells. RT-PCR and western blot assays were performed to detect expression of altered genes and proteins, and the MTT assay was used to detect cell viability. Our data showed that beta-catenin mRNA and protein expression levels were effectively knocked down by beta-catenin siRNA and subsequently, tumor cell proliferation was significantly suppressed. Flow cytometry assay showed that tumor cells were arrested at the G0/G1 phase of the cell cycles. Molecularly, expression of Smad3, p-caspase-3, and Grp78 protein were upregulated after 72 h of beta-catenin siRNA transfection, whereas expression of TERT, caspase-3, XIAP, MMP-2, MMP-9, VEGF-A, VEGF-c, and bFGF protein were reduced. However, there was no change between the expression of STAT3 and the HSP27 protein following transfection. The results from the current study demonstrated the importance of the Wnt/beta-catenin signaling pathway in regulation of gene expression in HCC. Further studies are required to investigate the role of this pathway in HCC development and targeting of this pathway to control HCC.
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Carcinoma Hepatocelular/genética , Regulación Neoplásica de la Expresión Génica/genética , Terapia Genética/métodos , Neoplasias Hepáticas/genética , Neovascularización Patológica/genética , beta Catenina/genética , Western Blotting , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Proliferación Celular , Separación Celular , Chaperón BiP del Retículo Endoplásmico , Citometría de Flujo , Expresión Génica , Humanos , Neoplasias Hepáticas/metabolismo , ARN Interferente Pequeño , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/genética , Transfección , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , beta Catenina/metabolismoRESUMEN
BACKGROUND: Experiments have reported that granulocyte colony stimulating factor (G-CSF) can mobilize stem cells. However, few studies have examined the effect of G-CSF on bone marrow mononuclear cell (BMMC) mobilization, in particular regarding their capability to home to acutely injured liver. AIMS: The aim of this study was to evaluate the effort of G-CSF on BMMC homing to the liver following chemically-induced hepatic failure. METHODS: BMMC were isolated from mice, pre-labeled with PKH26 and infused into the mice in which hepatic injury had been induced followed by administration of G-CSF or vehicle. Livers were studied by fluorescent microscopy after transplantation of pre-labeled BMMC. RESULTS: PKH26 labeled cells were found in liver tissue at 102 ± 10 cells/high power field in the BMMC+G-CSF group and 30 ± 5 cells/high power field in the BMMC group, but none in the G-CSF group and the control group (P < 0.05). In the former two groups the majority of PKH26 labeled cells colocalized with proliferative cell nuclear antigen (PCNA). The number of PCNA positive cells in the BMMC+G-CSF group was 20 ± 4 cells/high power field, while in the BMMC group it was 14 ± 2 cells/high power field, in the G-CSF group 12 ± 2 cells/high power field, and 8 ± 1 cells/high power field in the control group. Moreover, albumin expression was increased in the BMMC+G-CSF treated group (149 ± 7/high power field) relative to the BMMC group (48 ± 6/high power field), the G-CSF group (44 ± 5/high power field) and the vehicle group (30 ± 6/high power field), with the former three groups showing elevated levels as compared to vehicle control (30 ± 6) (P < 0.05). CONCLUSION: Transplanted BMMC may home to injured liver, which appears to be enhanced by G-CSF administration.
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Células de la Médula Ósea/efectos de los fármacos , Trasplante de Médula Ósea , Factor Estimulante de Colonias de Granulocitos/farmacología , Movilización de Célula Madre Hematopoyética/métodos , Células Madre Hematopoyéticas/efectos de los fármacos , Fallo Hepático Agudo/tratamiento farmacológico , Albúminas/metabolismo , Animales , Biopsia , Células de la Médula Ósea/citología , Tetracloruro de Carbono/toxicidad , Movimiento Celular/efectos de los fármacos , Células , Modelos Animales de Enfermedad , Citometría de Flujo , Colorantes Fluorescentes , Células Madre Hematopoyéticas/citología , Inmunohistoquímica , Hígado/citología , Fallo Hepático Agudo/inducido químicamente , Fallo Hepático Agudo/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Compuestos OrgánicosRESUMEN
BACKGROUND/AIMS: To investigate in patients the relationship between hepatocellular carcinoma and expression levels of cytochrome P450 IIE1 and glutathione S-transferaseP1 METHODOLOGY: Peripheral blood was collected from 65 patients with hepatocellular carcinoma, and 65 healthy controls. Real-time polymerase chain reaction was used in this research. RESULTS: The average mRNA levels of CYPIIE1 in HCC and healthy controls are 11.09% and 2.13%, respectively, while the average mRNA levels of GSTP1 in HCC and healthy controls are 0.61% and 2.34%, respectively. The mRNA level of CYPIIE1 was higher, and that of GSTP1 was lower, in patients with HCC compared to healthy controls. The difference of the mRNA levels of two enzymes in HCC has a statistical significance (p < 0.001). CONCLUSIONS: In this group of HCC patients and healthy controls, we found that patients with HCC tended to have higher expression of CYPIIE1 and lower expression of GSTP1. Our study indicate that CYPIIE1 is maybe the ruinous gene that results in an increased incidence of HCC, while GSTP1 is maybe the protection gene for hepatic cells during the whole course of metabolization.
Asunto(s)
Carcinoma Hepatocelular/enzimología , Citocromo P-450 CYP2E1/genética , Gutatión-S-Transferasa pi/genética , Neoplasias Hepáticas/enzimología , ARN Mensajero/análisis , Anciano , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
AIM: To evaluate the number of bone marrow mononuclear cells (BMMC) that are migrated to the liver following transplantation of murine BMMC into mice with acute liver injury. METHODS: BMMC were isolated from the bone marrow of mice in a lymphocyte separation medium and then labeled with PKH26. The labeled cells were subsequently infused into the caudal veins of BALB/c mice with hepatic injury induced by carbon tetrachloride and 2-acetylaminofluorene. Mice in experimental group were treated with stromal cell-derived factor-1 (SDF-1) which was injected intraperitoneally after transplantation of BMMC. Mice in control group were injected intraperitoneally with 0.1 mL of saline (0.9% NaCl) after transplantation of BMMC. After 2 wk, migration of the cells in experimental group was studied by fluorescence microscopy. The expression of proliferating cell nuclear antigen and albumin was quantified with manual methods in both groups. The serum transaminase levels at different time points were compared between the two groups. RESULTS: The labeled "cells" were found in the portal region and central veins of hepatic lobules. The PKH26-labeled cells appeared at an average frequency of 108 +/- 8/high power field in the experiment group and 65 +/- 8/high power field in the control group (P < 0.05). The total number of positive cells was 29 +/- 7/high power field in the experimental group and 13 +/- 2/high power field in the control group. The albumin expression level was also higher in the experimental group than in the control group (29 +/- 7 vs 13 +/- 2, P < 0.05). The total number of crossing points was 156 +/- 5/high power field in the experimental group and 53 +/- 5/high power field in the control group (P < 0.05). The serum alanine aminotransferase levels in experimental and control groups were measured at different time points (120 +/- 40 vs 118.50 +/- 1.75, P > 0.05; 80.60 +/- 6.50 vs 101.08 +/- 5.67, P < 0.05; 50.74 +/- 5.38 vs 80.47 +/- 4.62, P < 0.05; 30.54 +/- 2.70 vs 60.72 +/- 4.37, P < 0.05; 30.77 +/- 5.36 vs 40.47 +/- 6.50, P < 0.05). At the same time, the serum aspartate aminotransferase levels were measured in experimental and control groups at different time points (122.55 +/- 1.46 vs 120.70 +/- 4.22, P > 0.05; 54.26 +/- 6.50 vs 98.70 +/- 8.20, P < 0.05; 39.47 +/- 5.39 vs 78.34 +/- 4.50, P < 0.05; 28.94 +/- 2.70 vs 56.44 +/- 4.28, P < 0.05; 30.77 +/- 5.45 vs 42.50 +/- 6.28, P < 0.05). CONCLUSION: SDF-1 can promote the migration of BMMC to the liver of mice with acute liver failure.
Asunto(s)
Células de la Médula Ósea/fisiología , Trasplante de Médula Ósea , Movimiento Celular/fisiología , Quimiocina CXCL12/metabolismo , Leucocitos Mononucleares/fisiología , Fallo Hepático Agudo/metabolismo , Trasplante de Células Madre Mesenquimatosas , Albúminas/metabolismo , Animales , Células de la Médula Ósea/citología , Quimiocina CXCL12/genética , Modelos Animales de Enfermedad , Humanos , Leucocitos Mononucleares/citología , Hígado/citología , Hígado/metabolismo , Hígado/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Antígeno Nuclear de Célula en Proliferación/metabolismo , Transaminasas/sangreRESUMEN
BACKGROUND: Alcohol abuse and dependence are major factors in the pathogenesis of alcoholic liver disease (ALD). Alcohol abuse is becoming an increasingly severe problem among the Han, Mongol, and Korean nationalities in northeast China. This study aimed to investigate the relationship between ALD and the genetic polymorphism and expression levels of two enzymes, cytochrome P450IIE1 (CYPIIE1) and glutathione S-transferase P1 (GSTP1) in patients of three nationalities. METHODS: Peripheral blood was collected from 353 Chinese patients with ALD, 300 alcohol dependent patients without liver disease (alcoholic), and 360 healthy controls. Each group included patients from the Han, Mongol and Korean nationalities. Real-time polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) were used. RESULTS: Regardless of nationality, patients who carried the rare CYPIIE1 C2 and GSTP1 Val alleles were at higher risk of ALD. The frequency of C2 and Val in patients with ALD was respectively 50.00% and 26.98% in the Han, 31.36% and 22.87% in the Mongol, and 45.87% and 22.02% in the Korean nationality. No significant differences were seen in the frequency of either C2 or Val alleles in ALD patients among the three nationalities. In each nationality, the frequency of both C2 and Val alleles was significantly higher in ALD compared to alcoholic and healthy controls. Except for nationality, the average mRNA levels of CYPIIE1 in ALD patients and healthy controls were 10.05% and 2.21%, respectively. The average mRNA levels of GSTP1 in ALD patients and healthy controls were 0.53% and 2.12%, respectively. The mRNA level of CYPIIE1 was higher, and that of GSTP1 was lower in patients with ALD compared to the controls. CONCLUSIONS: Except for nationality, patients with ALD in this series tended to have a higher mRNA expression of CYPIIE1 and to carry the C2 allele, and tended to have a lower mRNA expression of GSTP1 and to carry the Val allele. There is a causal relationship between the polymorphic alleles, which leads to different mRNA levels and the development of ALD.