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1.
Anticancer Res ; 40(5): 2627-2635, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32366407

RESUMEN

BACKGROUND/AIM: Head and neck squamous cell carcinoma (HNSCC) is an aggressive head and neck malignancy. The aim of this study was to elucidate the role of periostin (POSTN) in the epithelial-to-mesenchymal transition (EMT) process mediating the acquisition of radioresistance in HNSCC. MATERIALS AND METHODS: The expression levels of EMT hallmark genes including POSTN and Erk/Akt signaling pathways were compared between radiosensitive and radioresistant HNSCC cells. RESULTS: POSTN mRNA expression was higher in radioresistant HNSCC cells, and silencing POSTN significantly impaired their invasiveness under the effect of EMT process represented by up-regulation of mesenchymal markers and down-regulation of an epithelial marker. Expression levels of Erk and Akt were higher in radioresistant cells. CONCLUSION: POSTN in association with the Erk and Akt signaling pathways was up-regulated during the EMT process, leading to the conversion of radiosensitive to radioresistant HNSCC cells. POSTN may be a key marker for predicting the radioresistance and therapeutic target of HNSCC.


Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Transición Epitelial-Mesenquimal , Neoplasias de Cabeza y Cuello/metabolismo , Neoplasias de Cabeza y Cuello/patología , Tolerancia a Radiación , Biomarcadores de Tumor/metabolismo , Línea Celular Tumoral , Regulación hacia Abajo , Epitelio/metabolismo , Epitelio/patología , Regulación Neoplásica de la Expresión Génica , Neoplasias de Cabeza y Cuello/genética , Humanos , Mesodermo/patología , Invasividad Neoplásica , Tolerancia a Radiación/genética , Transducción de Señal/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/patología
2.
Anticancer Res ; 40(5): 2645-2655, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32366409

RESUMEN

BACKGROUND/AIM: Two-thirds of head and neck squamous cell carcinoma (HNSCC) patients present with locally advanced (LA) stages and have a poor survival rate. The aim of this study was to investigate the roles of the long non-coding RNAs MALAT1 on radiation and cisplatin sensitivity of HNSCC cells. MATERIALS AND METHODS: Clonogenic, cell viability, and apoptosis assays were performed in cells following MALAT1 knockdown using CRISPR/Cas9 system. RESULTS: MALAT1 was overexpressed in HNSCC cell lines as compared to a non-tumorigenic cell line. The number of colonies formed after radiation was significantly reduced in MALAT1 knockdown cells. The IC50 value of cisplatin in MALAT1 knockdown cells was lower than that of the control cells. MALAT1 knockdown resulted in cell cycle arrest at G2/M phase, DNA damage and apoptotic cell death. CONCLUSION: MALAT1 knockdown enhanced the sensitivity of HNSCC cells to radiation and cisplatin partly through the induction of G2/M cell cycle arrest resulting in DNA damage and apoptosis.


Asunto(s)
Cisplatino/uso terapéutico , ARN Largo no Codificante/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/terapia , Apoptosis/genética , Puntos de Control del Ciclo Celular/genética , Línea Celular Tumoral , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , ARN Largo no Codificante/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/tratamiento farmacológico , Carcinoma de Células Escamosas de Cabeza y Cuello/radioterapia
3.
Anticancer Res ; 40(5): 2675-2685, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32366412

RESUMEN

BACKGROUND/AIM: To evaluate the anti-cancer mechanism of N-Farnesyl-norcantharimide (NC15). MATERIALS AND METHODS: The viability of NC15-treated human leukemic Jurkat T (JKT) cells was assessed using the Kit-8 cell counting method. Flow cytometry analysis, human apoptosis antibody array assay, and whole genome sequencing were adopted to investigate the mechanism underlying the anti-cancer activity of NC15 in JKT cells. RESULTS: The growth inhibition rates of NC15 in JKT cells were about 80% and 95% after treatment with 8 µmol/l NC15 for 24 and 48 h, respectively. The percentages of NC15-treated JKT cells in the sub-G1 phase at 24 and 48 h were 22.0% and 34.3%, respectively, in contrast to the 1.5% in the control. Next-generation sequencing showed that many tumor suppressor genes (TSG) were up-regulated, while many genes associated with steroid biosynthesis, metabolic pathways, and fatty acid metabolism were down-regulated. CONCLUSION: NC15 can reduce the cell viability and increase the percentage of JKT cells in the sub-G1 phase by up-regulating TSG and related genes, and down-regulating the genes for steroid biosynthesis, metabolic pathways and fatty acid metabolism, instead of through apoptosis.


Asunto(s)
Cantaridina/análogos & derivados , Regulación hacia Abajo/efectos de los fármacos , Ácidos Grasos/metabolismo , Genes Supresores de Tumor , Redes y Vías Metabólicas/genética , Esteroides/biosíntesis , Linfocitos T/citología , Regulación hacia Arriba/efectos de los fármacos , Apoptosis/efectos de los fármacos , Apoptosis/genética , Vías Biosintéticas/efectos de los fármacos , Vías Biosintéticas/genética , Cantaridina/química , Cantaridina/farmacología , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Proliferación Celular/efectos de los fármacos , ADN de Neoplasias/metabolismo , Regulación hacia Abajo/genética , Humanos , Células Jurkat , Redes y Vías Metabólicas/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Regulación hacia Arriba/genética
4.
Med Sci Monit ; 26: e922148, 2020 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-32275644

RESUMEN

BACKGROUND Sorafenib, which is a multitargeted kinase inhibitor, has shown some antitumor effects in patients with non-small cell lung cancer (NSCLC). However, the potential target of sorafenib's antitumor activity is largely unknown. Moreover, definitive predictive biomarkers of benefit have rarely been reported. MATERIAL AND METHODS The alteration in inhibitor of differentiation 1 (ID1) expression in NSCLC cells with sorafenib treatment was detected by western blotting. The sensitivity of NSCLC cells to sorafenib was observed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay. Loss-of-function and gain-of-function experiments were performed to observe the role of ID1 expression in epithelial to mesenchymal transition (EMT) progression. RESULTS Initially, we observed that ID1 was downregulated in NSCLC cells treated with sorafenib. The response of NSCLC cells to sorafenib was inhibited by the transfection of small interfering RNAs (siRNAs) targeting ID1. In contrast, the transfection of ID1-overexpressing plasmids improved the response of NSCLC cells to sorafenib. Further experiments indicated that ID1 is expressed at high levels in epithelial H460 cells and expressed at low levels in mesenchymal H358 cells. Loss-of-function and gain-of-function experiments suggested that ID1 negatively regulates EMT in NSCLC. CONCLUSIONS The expression of ID1 is dose-dependently inhibited by sorafenib, and the overexpression of ID1 contributes to the antitumor activity of sorafenib by suppressing EMT development. Our results indicate that ID1 might be a potential target for the antitumor activity of sorafenib in NSCLC and that targeting ID1 is a feasible strategy to improve the sensitivity of NSCLC cells to sorafenib.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/patología , Transición Epitelial-Mesenquimal/efectos de los fármacos , Proteína 1 Inhibidora de la Diferenciación/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Sorafenib/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Línea Celular Tumoral , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Interferencia de ARN , ARN Interferente Pequeño
5.
Med Sci Monit ; 26: e922092, 2020 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-32277808

RESUMEN

BACKGROUND Despite scientific advancement in radiotherapy and chemotherapy, the 5-year survival rate of lung cancer patients is around 15%. The present study explored the anticancer potential of betulinic acid nanoparticles against lung cancer cells. MATERIAL AND METHODS The proliferative changes in lung cancer cells by betulinic acid nanoparticles were measured by MTT assay. Cell cycle analysis was performed by flow cytometry using propidium iodide stain. Transwell and wound healing assay were used for determination of HKULC2 cell metastatic potential. RESULTS The betulinic acid nanoparticle treatment significantly (P<0.05) reduced proliferation of HKULC2, H1299, and H23 cells. The proliferation of HKULC2, H1299, and H23 cells was reduced to 33%, 28% and 24%, respectively on treatment with 10 µM of betulinic acid nanoparticles. The results from flow cytometry showed that betulinic acid nanoparticle exposure lead to cell cycle arrest in G1 phase in HKULC2 cells. Treatment with betulinic acid nanoparticles markedly decreased migration potential of HKULC2 cells. The invasive ability of HKULC2 cells was also suppressed markedly on exposure to betulinic acid nanoparticles. Western blotting of HKULC2 cells showed that betulinic acid nanoparticles promoted the expression of p21 and p53 and downregulated CD133, ALDH, BCL2, MCL1, and c-Myc expression. Betulinic acid nanoparticles reduced the expression of ABCG1 protein markedly. CONCLUSIONS The present study demonstrated that betulinic acid nanoparticles inhibit proliferation, metastatic ability, and arrest cell cycle in lung cancer cells through downregulation of ABCG1 oncogene expression. Therefore, betulinic acid nanoparticles may be used as therapeutic agent for the treatment of lung cancer.


Asunto(s)
Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1/genética , Antineoplásicos Fitogénicos/farmacología , Neoplasias Pulmonares/patología , Triterpenos/farmacología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Nanopartículas
6.
Eur Rev Med Pharmacol Sci ; 24(6): 3426-3432, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-32271462

RESUMEN

The outbreak of Sars-CoV-2 (COVID-19) poses serious challenges to people's health worldwide. The management of the disease is mostly supportive, and respiratory failure from acute respiratory distress syndrome is the leading cause of death in a significant proportion of affected patients. Preliminary data point out that dramatic increase in IL-6 and subsequent cytokine release syndrome may account for the development of fatal interstitial pneumonia. Inhibition of IL-6 by blocking its specific receptor with monoclonal antibodies has been advocated as a promising attempt. Here we assess the potential utility of myo-Inositol, a polyol already in use for treating the newborn Respiratory Distress Syndrome, in downregulating the inflammatory response upon Sars-CoV-2 infection. Myo-Inositol proved to reduce IL-6 levels in a number of conditions and to mitigate the inflammatory cascade, while being devoid of any significant side effects. It is tempting to speculate that inositol could be beneficial in managing the most dreadful effects of Sars-CoV-2 infection.


Asunto(s)
Betacoronavirus/efectos de los fármacos , Infecciones por Coronavirus/tratamiento farmacológico , Síndrome de Liberación de Citoquinas/etiología , Inositol/uso terapéutico , Neumonía Viral/tratamiento farmacológico , Infecciones por Coronavirus/complicaciones , Infecciones por Coronavirus/fisiopatología , Regulación hacia Abajo , Humanos , Interleucina-6/metabolismo , Neoplasias Pulmonares/metabolismo , Pandemias , Neumonía Viral/complicaciones , Neumonía Viral/fisiopatología
7.
Autoimmun Rev ; 19(6): 102536, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32251718
9.
Life Sci ; 251: 117634, 2020 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-32251632

RESUMEN

Neuregulin-1ß (NRG-1) is a membrane-bound or secreted growth and differentiation factor that mediates its action by binding to ErbB receptors. Circulating levels of NRG-1 are characterized by large inter-individual variability with the range of absolute values covering two orders of magnitude, from hundreds to tens of thousands of picograms per milliliter of blood. NRG-1 signaling via ErbB receptors contributes to the cell survival and downregulation of the inflammatory response. A higher level of circulating NRG-1 may indicate increased shedding of membrane-bound NRG-1, which in turn can contribute to better protection against cardiovascular stress or injury. However, it is unknown whether circulating NRG-1 can induce activation of ErbB receptors. In the current study, we performed an analysis of circulating NRG-1 functional activity using a cell-based ELISA measuring phosphorylation of ErbB3 induced by blood plasma obtained from healthy donors. We found high levels of ErbB3 activating activity in human plasma. No correlations were found between the levels of circulating NRG-1 and plasma ErbB3 activating activity. To determine the direct effect of circulating NRG-1, we incubated plasma with neutralizing antibody, which prevented the stimulatory effect of recombinant NRG-1 on activation of ErbB3. No effect of the neutralizing antibody was found on plasma-induced phosphorylation of ErbB3. We also found that a significant portion of circulating NRG-1 is comprised of full-length NRG-1 associated with large extracellular vesicles. Our results demonstrate that circulating NRG-1 does not contribute to plasma-induced ErbB3 activating activity and emphasizes the importance of functional testing of NRG-1 proteins in biological samples.


Asunto(s)
Supervivencia Celular/fisiología , Neurregulina-1/metabolismo , Receptor ErbB-3/metabolismo , Adulto , Anciano , Anticuerpos Neutralizantes/inmunología , Regulación hacia Abajo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fosforilación
10.
Gene ; 747: 144653, 2020 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-32259630

RESUMEN

Diabetic retinopathy (DR) is a frequent complication of diabetes and it can lead to visual impairment and blindness. However, the mechanism of their regulation remains little known. circRNAs can function as crucial competing endogenous RNA, which can sponge corresponding miRNAs and affect mRNA expression in various diseases, including DR. In our current research, we observed that circRNA_0084043 was elevated in high glucose (HG)-incubated ARPE-19 cells. Then, we focused on whether and how circRNA_0084043 participated in retinal vascular dysfunction under conditions diabetes. Apoptosis, inflammation and oxidative stress are hallmark of DR progression. This work was aimed to investigate the signaling mechanisms of circRNA_0084043 in these pathogenesis of DR. We discovered loss of circRNA_0084043 significantly increased cell survival and repressed HG-triggered apoptosis. In addition, knockdown of circRNA_0084043 remarkably reduced oxidative stress as evidenced by the down-regulated malondialdehyde (MDA) content, enhanced activities of Super Oxide Dismutase (SOD) and Glutathione peroxidase (GSH-PX). Addition, silence of circRNA_0084043 effectively restrained HG-stimulated inflammation as proved by repressing inflammatory cytokines Tumor Necrosis Factor α (TNF-α), Interleukin 6 (IL-6) and Cox-2 in ARPE-19 cells. Subsequently, we successfully predicted that miR-140-3p was a downstream target of circRNA_0084043, which could be negatively regulated by circRNA_0084043. Mechanistically, loss of miR-140-3p abrogated the beneficial effects of circRNA_0084043 siRNA on ARPE-19 cells. Transforming Growth Factor alpha (TGFA) can exhibit a role in multiple diseases. Taken these together, these data demonstrated that loss of circRNA_0084043 depressed HG-induced damage via sponging miR-140-3p and regulating TGFA.


Asunto(s)
Retinopatía Diabética/genética , Progresión de la Enfermedad , Células Epiteliales/metabolismo , Regulación de la Expresión Génica , MicroARNs/metabolismo , ARN Circular/genética , Epitelio Pigmentado de la Retina/patología , Factor de Crecimiento Transformador alfa/genética , Apoptosis/efectos de los fármacos , Secuencia de Bases , Línea Celular , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Regulación de la Expresión Génica/efectos de los fármacos , Glucosa/toxicidad , Humanos , Inflamación/patología , MicroARNs/genética , Modelos Biológicos , Estrés Oxidativo/efectos de los fármacos , ARN Circular/metabolismo , Factor de Crecimiento Transformador alfa/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética
11.
Gene ; 747: 144654, 2020 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-32259632

RESUMEN

Diabetic retinopathy (DR)is a common diabetes complication, resulting in the loss of vision. circRNAshave been reported to serve as ceRNA via targeting corresponding miRNAs and modulating mRNA expression in various diseases. Recently, increasing reports has indicated circRNAs can exert a significant role inDR progression. However, the expression and mechanism of hsa_circ_0041795 in human retinal pigment epithelial cells ARPE-19 treated by high glucose remains poorly known. Hence, we aimed to work figure out the effect of hsa_circ_0041795 in high glucose (HG)-induced ARPE-19 cell damage and study its molecular mechanisms. In our current research, we found that hsa_circ_0041795 was obviously up-regulated in HG-treated ARPE-19 cells. High dose of glucose greatly depressed ARPE-19 cell survival and contributed to cell apoptosis. In addition, we observed that loss of hsa_circ_0041795 enhanced cell proliferation and inhibit ARPE-19 cell apoptosis, after HG incubation. Furthermore, data of ELISA indicated that hsa_circ_0041795 siRNA significantly restrained inflammatory factors expression, such as TNF-α, IL-1ß and IL-6 in ARPE-19 cells treated with HG. miR-646 has been recognized in multiple diseases and currently, we predicted that miR-646 acted as a target of hsa_circ_0041795. Moreover, we found that miR-646 inhibitors dramatically reversed the effect of hsa_circ_0041795 siRNA on ARPE-19 cells. Additionally, a dual-luciferase reporter assay proved that VEGFC was a direct target of miR-646. Our results demonstrated that hsa_circ_0041795 might exhibit a novel therapeutic potential in the treatment of DR.


Asunto(s)
Células Epiteliales/metabolismo , Células Epiteliales/patología , Glucosa/toxicidad , MicroARNs/metabolismo , ARN Circular/metabolismo , Epitelio Pigmentado de la Retina/lesiones , Factor C de Crecimiento Endotelial Vascular/metabolismo , Apoptosis/efectos de los fármacos , Apoptosis/genética , Secuencia de Bases , Línea Celular , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Células Epiteliales/efectos de los fármacos , Humanos , Inflamación/genética , Inflamación/patología , MicroARNs/genética , ARN Circular/genética , ARN Interferente Pequeño/metabolismo
12.
Med Sci Monit ; 26: e921510, 2020 Apr 02.
Artículo en Inglés | MEDLINE | ID: mdl-32238796

RESUMEN

BACKGROUND SOX7 exerts a repressing effect against tumors and imposes vital influences on malignancies. Our research discussed the importance of SOX7 in breast cancer prognoses. MATERIAL AND METHODS SOX7 mRNA expression in breast cancer tissues samples and matched adjacent normal controls of breast cancer patients was measured by quantitative real-time-polymerase chain reaction (qRT-PCR). The relationship of SOX7 with clinicopathological characteristics were analyzed via chi-square test. The association of SOX7 levels with clinical outcomes was evaluated adopting the Kaplan-Meier method and multivariate Cox proportional hazards regression model. RESULTS SOX7 mRNA degree of expression exhibited a declining tendency in breast cancer tissue compared to paired bordering normal tissue specimens (P<0.001). In addition, the reduced SOX7 degree of expression had a strong correlation to larger cancer mass dimension (P=0.006) and lymph node metastasis (P=0.001). Survival analysis revealed that the overall survival (OS) time was much shorter among cases harboring low SOX7 degree of expression compared to high degree of expression (P=0.005). Moreover, SOX7 expression alone could predict OS among breast cancer patients (hazard ratio=3.956, 95% confidence interval=1.330-11.772, P=0.013). CONCLUSIONS SOX7 expression was downregulated in breast cancer tissues, and it could function as a useful prognostic marker in breast cancer.


Asunto(s)
Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Factores de Transcripción SOXF/metabolismo , Biomarcadores de Tumor/genética , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/patología , Regulación hacia Abajo , Femenino , Perfilación de la Expresión Génica , Humanos , Metástasis Linfática , Pronóstico , Modelos de Riesgos Proporcionales , Factores de Transcripción SOXF/genética , Análisis de Supervivencia
13.
Med Sci Monit ; 26: e920628, 2020 Apr 03.
Artículo en Inglés | MEDLINE | ID: mdl-32242546

RESUMEN

BACKGROUND Due to its remarkable effect in controlling glycometabolism, relatively simple operation, and low risk of complications, sleeve gastrectomy (SG) has become the preferred surgical treatment for type II diabetes mellitus. Increased blood glucose in the body can cause damage to functional cells. MATERIAL AND METHODS Long non-coding RNA SNHG5 expression and TGR5 in serum were analyzed by real-time PCR. A diabetic cell model was established by culturing normal human intestinal epithelial cells NCM460 and DLD-1 with high-glucose and high-fat medium. CCK-8 assay, TUNEL assay, and flow cytometry were used to assess cell growth and apoptosis, respectively. The secretion of lactate dehydrogenase (LDH) was detected using the LDH Cytotoxicity Kit. lncRNA SNHG5 was downregulated by siRNA. The changes in expression of SNHG5, TGR5, Akt, p65, and Bcl-2 were analyzed by real-time PCR assay or Western blot. RESULTS In 40 type II diabetes patients who underwent sleeve gastrectomy, the expression of SNHG5 decreased and the expression of TGR5 increased compared with that before the operation. After high-glucose and high-fat culture, cell growth was inhibited and cell apoptosis increased significantly. The expression of SNHG5 was increased and TGR5 was decreased with high-glucose and high-fat culture. However, high glucose and high fat showed an opposite trend for cell growth, apoptosis, and LDH release under inhibition of SNHG5. The expression levels of TGR5 and Akt, p65, and Bcl-2 were also returned to normal by SNHG5 inhibition. CONCLUSIONS By downregulating expression of the SNHG5 gene and then altering expression of the TGR5 gene, the damage to colorectal cells induced by high glucose was alleviated. This may be one of the mechanisms underlying the effect of sleeve gastric surgery in treatment of diabetes mellitus.


Asunto(s)
Glucemia/metabolismo , Diabetes Mellitus Tipo 2/cirugía , Gastrectomía/métodos , Receptores Acoplados a Proteínas G/metabolismo , Apoptosis , Proliferación Celular , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/metabolismo , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Humanos , Secreción de Insulina , Obesidad/complicaciones , Obesidad/metabolismo , Obesidad/cirugía , ARN Largo no Codificante , Receptores Acoplados a Proteínas G/agonistas
14.
Biol Res ; 53(1): 17, 2020 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-32312329

RESUMEN

BACKGROUND: Inflammation and apoptosis are considered to be two main factors affecting ischemic brain injury and the subsequent reperfusion damage. MiR-19a-3p has been reported to be a possible novel biomarker in ischemic stroke. However, the function and molecular mechanisms of miR-19a-3p remain unclear in cerebral ischemia/reperfusion (I/R) injury. METHODS: The I/R injury model was established in vivo by middle cerebral artery occlusion/reperfusion (MCAO/R) in rats and in vitro by oxygen-glucose deprivation and reperfusion (OGD/R) induced SH-SY5Y cells. The expression of miR-19a-3p was determined by reverse transcription quantitative PCR. The infarction volumes, Neurological deficit scores, apoptosis, cell viability, pro-inflammatory cytokines and apoptosis were evaluated using Longa score, Bederson score, TTC, TUNEL staining, CCK-8, ELISA, flow cytometry assays. Luciferase reporter assay was utilized to validate the target gene of miR-19a-3p. RESULTS: We first found miR-19a-3p was significantly up-regulated in rat I/R brain tissues and OGD/R induced SH-SY5Y cells. Using the in vivo and in vitro I/R injury model, we further demonstrated that miR-19a-3p inhibitor exerted protective role against injury to cerebral I/R, which was reflected by reduced infarct volume, improved neurological outcomes, increased cell viability, inhibited inflammation and apoptosis. Mechanistically, miR-19a-3p binds to 3'UTR region of IGFBP3 mRNA. Inhibition of miR-19a-3p caused the increased expression of IGFBP3 in OGD/R induced SH-SY5Y cells. Furthermore, we showed that IGFBP3 overexpression imitated, while knockdown reversed the protective effects of miR-19a-3p inhibitor against OGD/R-induced injury. CONCLUSIONS: In summary, our findings showed miR-19a-3p regulated I/R-induced inflammation and apoptosis through targeting IGFBP3, which might provide a potential therapeutic target for cerebral I/R injury.


Asunto(s)
Isquemia Encefálica/genética , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , MicroARNs/genética , Oxígeno/metabolismo , Daño por Reperfusión/genética , Animales , Apoptosis , Biomarcadores/metabolismo , Isquemia Encefálica/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Glucosa/deficiencia , Masculino , Neuronas/metabolismo , Neuroprotección , ARN Mensajero/genética , Distribución Aleatoria , Ratas , Daño por Reperfusión/metabolismo , Accidente Cerebrovascular , Regulación hacia Arriba
15.
Toxicon ; 180: 11-17, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32259553

RESUMEN

Ochratoxin A (OTA) is a toxic metabolite produced by Aspergillus and Penicillium fungus. OTA found in the human and animal tissues can contaminate many foods that we daily consume in our lives. It accumulates especially in kidney. Although OTA is known to cause cell cycle arrest, the molecular mechanisms underlying this effect have not been fully understood, yet. We aimed to investigate the molecular details of OTA induced inhibitory response in G1 - G1/S phase of cell cycle and also the regulatory role of p53 in OTA mediated cell cycle arrest in human proximal tubule epithelial cells, HK-2. For this purpose, Cyclin E1 and Cyclin D1 mRNA expressions and Cyclin D1, Cdk4 and Cdk2 protein expressions were evaluated in HK-2 cells transfected with either 50 nM control siRNA or p53 siRNA for 72 h in the absence or presence of OTA using RT-PCR and Western blot analyses, respectively. Our findings showed that mRNA expressions of Cyclin D1 and Cyclin E1 and protein expressions of Cyclin D1, Cdk4 and Cdk2 were inhibited in HK-2 cells treated with two different doses of OTA, 10 µM and 25 µM, for 24 h. However, the downregulation of p53 led to enhance OTA-mediated increase in mRNA expressions of Cyclin D1 and Cyclin E1 and protein expressions of Cyclin D1, Cdk4 and Cdk2 compared to control siRNA transfected HK-2 cells. Our findings strongly suggest that the cell cycle arresting effect of OTA also performs via a p53 mediated mechanism besides other possible mechanisms.


Asunto(s)
Puntos de Control del Ciclo Celular/efectos de los fármacos , Ocratoxinas/toxicidad , Proteínas de Ciclo Celular/metabolismo , Línea Celular , Ciclina D1 , Ciclina E , Regulación hacia Abajo , Fase G1/efectos de los fármacos , Humanos , Riñón , Proteínas Oncogénicas , Fase S/efectos de los fármacos , Pruebas de Toxicidad , Proteína p53 Supresora de Tumor/metabolismo
16.
Med Sci Monit ; 26: e921276, 2020 Apr 06.
Artículo en Inglés | MEDLINE | ID: mdl-32249762

RESUMEN

BACKGROUND Cartilage degeneration during osteoarthritis (OA) most adversely affects the quality of life by hindering the movement. The present study investigated the role of verbascoside in the protection of cartilage degeneration induced by osteoarthritis. MATERIAL AND METHODS The enzyme-linked immunosorbent (ELISA) and western blot assays were used for determination of inflammatory cytokine secretion in serum and cartilage tissues, respectively. RESULTS Treatment of the OA rats with verbascoside inhibited overproduction of interleukin (IL)-6, tumor necrosis factor (TNF)-alpha, and IL-1ß in serum as well as cartilage tissues. The expression of P2X7R and matrix metalloproteinase (MMP)-13 was much higher in the rats induced with OA. However, administration of verbascoside reversed the OA-induced upregulation of P2X7R and MMP-13 expression in the cartilage tissues. The OA-mediated increase in substance P (SP) and prostaglandin E2 (PGE2) expression was also reduced in the cartilage tissues by the verbascoside treatment. Western blot assay revealed that verbascoside treatment markedly decreased the activation of IkappaBalpha and NF-kappaB p65 in the OA rats. CONCLUSIONS Thus, verbascoside inhibited inflammatory cytokine secretion in the OA rats by targeting P2X7R expression, production of matrix metalloproteinase, PGE2 and downregulation of NF-kappaB signaling pathway. Therefore, verbascoside may be used as potent agent for osteoarthritis treatment.


Asunto(s)
Citocinas/metabolismo , Glucósidos/farmacología , Inmunosupresores/farmacología , FN-kappa B/metabolismo , Osteoartritis/prevención & control , Fenoles/farmacología , Animales , Cartílago Articular/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Antagonistas del Receptor Purinérgico P2X/farmacología , Calidad de Vida , Ratas
17.
J Cancer Res Clin Oncol ; 146(5): 1153-1167, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32189106

RESUMEN

BACKGROUND: Hepatoblastoma (HB) and pediatric hepatocellular carcinoma (HCC) are the most common malignant liver tumors in childhood. Both tumor types exhibit genetic and epigenetic alterations in the WNT/ß-catenin signaling pathway, which is a key regulator of liver progenitor cells in embryonic development. The tumors demonstrate a high rate of ß-catenin mutations and gene expression changes of several WNT antagonists. However, the role of the WNT inhibitory factor secreted frizzled-related protein 1 (SFRP1) has not been addressed in pediatric liver cancer so far. RESULTS: In our study, we investigated the gene expression level, DNA methylation status and functional relevance of SFRP1 in HB cell lines and in pediatric liver tumor patient samples. SFRP1 was downregulated due to DNA promoter methylation in all tested HB cell lines. Overexpression of SFRP1 in HB cell lines diminished tumor cell proliferation, colony formation and migration potential. In addition, the SFRP1-expressing HB cell lines showed reduced WNT/ß-catenin signaling pathway activity and decreased expression of WNT target genes. To evaluate the utility of SFRP1 as a biomarker in pediatric liver cancer, we determined the gene expression level and DNA methylation status of SFRP1 in 45 pediatric liver tumor patient samples. The correlation analysis of different clinical parameters and tumor characteristics revealed a significant correlation of reduced SFRP1 expression with the presence of mutant ß-catenin. The methylation status of SFRP1 was furthermore associated to a pediatric liver tumor type with HCC-like characteristics, TERT mutations and an older age at diagnosis. CONCLUSION: Altogether, our data demonstrate that the epigenetic suppression of the WNT/ß-catenin antagonist SFRP1 has an important impact on the malignant behavior of HB cells. Although SFRP1 methylation is a common event in HCC-like pediatric liver tumors, its potential as a prognostic or diagnostic biomarker needs to be further investigated.


Asunto(s)
Hepatoblastoma/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Neoplasias Hepáticas/genética , Proteínas de la Membrana/genética , Mutación , beta Catenina/genética , Anciano , Línea Celular Tumoral , Metilación de ADN , Regulación hacia Abajo , Femenino , Células Hep G2 , Hepatoblastoma/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intercelular/biosíntesis , Neoplasias Hepáticas/metabolismo , Masculino , Proteínas de la Membrana/biosíntesis , Persona de Mediana Edad , Regiones Promotoras Genéticas , Vía de Señalización Wnt , beta Catenina/metabolismo
18.
J Cancer Res Clin Oncol ; 146(5): 1125-1137, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32200459

RESUMEN

PURPOSE: Few studies reported about the potential of unphosphorylated heat shock protein 27 (HSP27) and phosphorylated heat shock protein 27 (pHSP27) as a predictor for survival and gemcitabine resistance in pancreatic ductal adenocarcinoma (PDAC). In this study, we analysed the expression patterns of pHSP27 and HSP27 in a patient population after surgery and correlated the immunohistochemical results with clinicopathological data and long-term outcome of the patients. METHODS: HSP27 and pHSP27 (Ser-15, Ser-78 and Ser-82) protein expression were analysed by immunohistochemistry using the immunoreactive score (IRS) from paraffin-embedded tissue of 106 patients with PDAC who underwent surgery. Immunohistochemical results were correlated with clinicopathological data, disease-free (DFS) and overall survival (OS). RESULTS: HSP27 expression was significantly lower in patients with a shorter OS (p = 0.006) and DFS (p < 0.0001). A higher HSP27 expression was associated with a better response to gemcitabine in the resected, non-metastasised patients group (p = 0.001). Furthermore, HSP27 was downregulated in patients suffering from metastases at time of surgery (p < 0.001) and in undifferentiated tumours (p = 0.007). In contrast, pHSP27-Ser15, -Ser78 and -Ser82 were not associated with any survival data of the study population. CONCLUSION: HSP27 seems to be a strong indicator for the prediction of OS and DFS. Moreover, HSP27 could play a role in the formation and migration of liver metastases of PDAC.


Asunto(s)
Carcinoma Ductal Pancreático/metabolismo , Proteínas de Choque Térmico/metabolismo , Chaperonas Moleculares/metabolismo , Neoplasias Pancreáticas/metabolismo , Anciano , Biomarcadores de Tumor/metabolismo , Carcinoma Ductal Pancreático/tratamiento farmacológico , Carcinoma Ductal Pancreático/patología , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacología , Regulación hacia Abajo , Resistencia a Antineoplásicos , Femenino , Proteínas de Choque Térmico/biosíntesis , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundario , Masculino , Persona de Mediana Edad , Chaperonas Moleculares/biosíntesis , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/patología , Fosforilación , Pronóstico , Tasa de Supervivencia
19.
Anticancer Res ; 40(3): 1375-1385, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32132034

RESUMEN

BACKGROUND/AIM: As metastasis accounts for most breast cancer (BC)-related deaths, identifying key players becomes research priority. Growth differentiation factor-15 (GDF15), a member of the transforming growth factor-ß superfamily, is affected by the actin cytoskeleton and has been associated with cancer. However, its exact role in BC cell invasiveness is vague. MATERIALS AND METHODS: GDF15 short-hairpin (shRNA)-mediated silencing was used to inhibit GDF15 expression in MCF-7 and MDA-MB-231 BC cells and gene expression of relevant focal adhesion (FA) genes, cell migration, invasion and tumor spheroid invasion were subsequently analyzed. RESULTS: GDF15 silencing promoted cell migration, cell invasion as well as tumor spheroid invasion and up-regulated urokinase plasminogen activator (uPA) and FA genes, integrin-linked kinase (ILK), LIM zinc finger domain containing 1 (LIMS1), α-parvin (PARVA), and RAS suppressor-1 (RSU1). Computational analysis of Cancer Genome Atlas BC dataset however, revealed no significant correlation between GDF15 expression and metastasis pointing towards a more complex molecular interplay between GDF15, actin cytoskeleton and FA-related genes which ultimately affects their expression pattern, in vivo. CONCLUSION: GDF15 suppresses BC cell invasion in vitro through down-regulation of FA genes but its role in BC is more complicated in vivo and warrants further investigation.


Asunto(s)
Neoplasias de la Mama/genética , Adhesiones Focales/genética , Factor 15 de Diferenciación de Crecimiento/genética , Neoplasias de la Mama/patología , Diferenciación Celular , Línea Celular Tumoral , Regulación hacia Abajo , Femenino , Humanos
20.
Gene ; 743: 144614, 2020 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-32222532

RESUMEN

Buffalo reproduction struggles with a high incidence of early embryonic mortality. Effective treatment and prevention strategies for this condition are not available due to lack of understanding of molecular pathways in early pregnancy of this species. In the present study, we have attempted to understand these molecular pathways by characterizing the endometrial transcriptomic profiles of pregnant buffalos during early pregnancy. For the transcriptome profiling, buffalo endometrial tissues of 29-36 days of pregnancy and of nonpregnant luteal phase were collected from the local slaughterhouse. We confirmed the status of pregnancy based on the crown vertebral length of the foetus. Total RNA was isolated and sequencing was performed using the Illumina nextseq platform. The raw reads were filtered and mapped to the Bos taurus UMD 3.1 reference genome assembly. An average of 24,597 genes was investigated for differential expression between the two groups. Transcriptome data identified a total of 450 differentially expressed genes (using a cut off value of log2 fold changes >2 and <-2) in early pregnancy in comparison to the nonpregnant group (Padj < 0.05). Among these, 270 genes were significantly upregulated and 180 genes were downregulated. The most impacted pathways were related to secretion, transport, ionic homeostasis, mitosis and negative regulation of viral processes. In conclusion, our study characterized a unique set of DEGs, during the early pregnancy of buffalo, which potentially modulate the endometrial environment to establish and maintain a successful pregnancy.


Asunto(s)
Búfalos/fisiología , Endometrio/metabolismo , Preñez/genética , ARN/metabolismo , Transcriptoma/fisiología , Animales , Regulación hacia Abajo , Femenino , Embarazo , ARN/genética , ARN/aislamiento & purificación , RNA-Seq , Regulación hacia Arriba
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