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Since December 2019, a new form of severe acute respiratory syndrome (SARS) from a novel strain of coronavirus (SARS coronavirus 2 [SARS-CoV-2]) has been spreading worldwide. The disease caused by SARS-CoV-2 was named Covid-19 and declared as a pandemic by the World Health Organization in March 2020. Clinical symptoms of Covid-19 range from common cold to more severe disease defined as pneumonia, hypoxia, and severe respiratory distress. In the next stage, disease can become more critical with respiratory failure, sepsis, septic shock, and/or multiorgan failure. Outcomes of Covid-19 indicate large gaps between the male-female and the young-elder groups. Several theories have been proposed to explain variations, such as gender, age, comorbidity, and genetic factors. It is likely that mixture of genetic and nongenetic factors interplays between virus and host genetics and determines the severity of disease outcome. In this review, we aimed to summarize current literature in terms of potential host genetic and epigenetic factors that associated with increased severity of Covid-19. Several studies indicated that the genetic variants of the SARS-CoV-2 entry mechanism-related (angiotensin-converting enzymes, transmembrane serine protease-2, furin) and host innate immune response-related genes (interferons [IFNs], interleukins, toll-like receptors), and human leukocyte antigen, ABO, 3p21.31, and 9q34.2 loci are critical host determinants related to Covid-19 severity. Epigenetic mechanisms also affect Covid-19 outcomes by regulating IFN signaling, angiotensin-converting enzyme-2, and immunity-related genes that particularly escape from X chromosome inactivation. Enhanced understanding of host genetic and epigenetic factors and viral interactions of SARS-CoV-2 is critical for improved prognostic tools and innovative therapeutics.
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COVID-19/epidemiología , COVID-19/genética , SARS-CoV-2/genética , Enzima Convertidora de Angiotensina 2/genética , COVID-19/enzimología , COVID-19/metabolismo , Epigénesis Genética , Epigenómica/métodos , Femenino , Furina/genética , Humanos , Inmunidad Innata/genética , Interferones/genética , Masculino , Pandemias , Peptidil-Dipeptidasa A/genética , Pronóstico , SARS-CoV-2/patogenicidad , Serina Endopeptidasas/genética , Índice de Severidad de la EnfermedadRESUMEN
PIKfyve is an evolutionarily conserved lipid and protein kinase enzyme that has pleiotropic cellular functions. The aim of the present study was to investigate the effects of phosphatidylinositol-3-phosphate 5-kinase (PIKfyve) inhibitor, YM201636, on nonsmall cell lung cancer (NSCLC) cells growth, tumorigenicity, and claudin (CLDN) expressions. Three NSCLC cell lines (Calu-1, H1299 and HCC827) were used to compare the effects of YM201636. Cytotoxic effects of YM201636 were analysed using XTT assay. Malignancy potential of cells assesses with wound healing and soft agar colony-forming assays. mRNA and protein expressions of claudins were analysed by qRT-PCR and immunofluorescence staining. Our results revealed that YM201636 inhibited the proliferation and malignancy potential of Calu-1, H1299, and HCC827 cells in a dose-dependent manner. After YM201636 treatment CLDN1, -3 and -5 expressions increased significantly in HCC827 cells. CLDN3 and -5 expressions also significantly increased in Calu1 cell line. YM201636 treatment significantly reduced the CLDN1 and increased the CLDN5 expression in H1299 cells. Immunofluorescence staining of CLDN1, -3 and -5 proteins showed a significant increase after YM201636 treatment. Besides, YM201636 induced EGFR mRNA expression in all NSCLC cell lines. Our results have shown that YM201636 inhibits tumorigenicity of NSCLC cells. Furthermore, estimated glomerular filtration rate (EGFR) pathway is important signalling involved in the regulation of claudins. Understanding the mechanisms of PIKfyve inhibitors may improve cancer treatment particularly for EGFR overactivated NSCLC.
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Lung cancer is one of the most commonly diagnosed cancers and it is associated with high rates of morbidity and mortality. Metastasis and relapse of the tumor depend on the survival and proliferation of lung cancer stem cells (LCSCs). The ability to identify CSCs may prevent recurrence and lead to more effective treatments. Sirtuins are a group of deacetylases that include seven variants (SIRT17), with sirtuin 1 (SIRT1) being the most intensively investigated. Evidence suggests that SIRT1 is both a tumorsuppressor gene and an oncogene. SIRT1 can deacetylate the tumorsuppressor protein p53 to decrease its activity. SIRT1 activators increase the deacetylation of p53, whereas SIRT1 inhibitors can stimulate p53 by inhibiting deacetylation. In the present study, CD44+ and CD133+enriched A549 (nonsmall cell lung cancer) cells collected using the CD44 and CD133 CSC surface markers by fluorescenceactivated cell sorting method were treated with SIRT1 inhibitors (tenovin6 and sirtinol) and SIRT1 activators (resveratrol and SRT1720), and their effects on apoptosis, as well as the mRNA and protein expression of SIRT1 and p53 were investigated. Of these agents, it was found that resveratrol increased p53 expression by 4.1fold, decreased SIRT1 expression by 0.2fold, and it was the most potent inducer of apoptosis.
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Antineoplásicos/farmacología , Benzamidas/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Neoplasias Pulmonares/tratamiento farmacológico , Resveratrol/farmacología , Sirtuina 1/metabolismo , Células A549 , Antígeno AC133/análisis , Antígeno AC133/metabolismo , Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Humanos , Receptores de Hialuranos/análisis , Receptores de Hialuranos/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Naftoles/farmacología , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Sirtuina 1/antagonistas & inhibidoresRESUMEN
PURPOSE: The purpose of this in vitro study was to evaluate the cytotoxic and apoptotic effects of seven new-generation all-ceramic materials for CAD/CAM (Lava Ultimate [LU], VITA Mark II [VM], InCoris TZI [IC], IPS e.max CAD [EM], VITA Suprinity [VS], Cerasmart [CS], IPS Empress CAD [EC]) and six provisional materials (Protemp 4 [PT], Telio CAD [TC], CAD-Temp [CT], Telio Lab [TL], Temdent Classic [TD], Telio CS C&B [TS]) on L929 mouse fibroblast cells. MATERIALS AND METHODS: 24 disc-shaped specimens (∅ = 5 mm, h = 2 mm) were prepared from each test material. Medium extracts were collected at the 1st, 3rd, and 7th days for each group and tested using the L929 cell line. Cytotoxicity was evaluated using XTT assay, and apoptosis was determined by Annexin-V/PI staining. Data were analyzed using one-way ANOVA, Tukey's multiple comparison tests at a significance level of p < 0.05. RESULTS: The cell viability results among all-ceramic material groups after the 1st and 7th days of incubation periods showed statistically significant differences (p < 0.05). There were significant differences within the ceramic groups in different incubation periods regarding apoptosis rate (p < 0.05). Throughout the entire test period, LU and VM from the CAD/CAM all-ceramic materials and PT and TC from the provisional restoration materials showed cell viability higher than 90%. EC and TD showed the lowest cell viability and highest apoptosis rates in their own groups. For the provisional materials, there were significant differences in cell viability and apoptosis rate in all the incubation periods for each material (p < 0.05). CONCLUSIONS: Although some new-generation CAD/CAM and provisional restoration materials display slight cytotoxicity values, the results are still within the reliable range, and they can safely be used in clinical conditions.
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Cerámica/uso terapéutico , Fibroblastos/efectos de los fármacos , Resinas Acrílicas/efectos adversos , Resinas Acrílicas/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Cerámica/efectos adversos , Resinas Compuestas/efectos adversos , Resinas Compuestas/uso terapéutico , Materiales Dentales/efectos adversos , Materiales Dentales/uso terapéutico , Porcelana Dental/efectos adversos , Porcelana Dental/uso terapéutico , Técnicas In Vitro , RatonesRESUMEN
OBJECTIVE: The aim of this study was to investigate the relationships between F216L (rs28942112), R496W (rs374603772), S127R (rs28942111), and D374Y (rs137852912) PCSK9 gain-of-function (GOF) mutations and primary dyslipidemia and serum lipid levels in patients with primary dyslipidemia. METHODS: In this case-control study, DNA was isolated from blood samples collected from patients diagnosed with primary dyslipidemia in cardiology outpatient clinic of Ege University (n=200) and healthy individuals (n=201). F216L, R496W, S127R, and D374Y GOF mutations in the PCSK9 gene were evaluated and genotyped according to the results of melting curve analysis performed in a real-time polymerase chain reaction (PCR) 480 instrument using specific primers for each mutation. RESULTS: There were statistically significant differences between the patient and individuals in control groups in the R496W and D374Y mutations (x2=10.742 p=0.005; x2=6.078 p=0.048, respectively). In addition, triglyceride levels in patients with primary dyslipidemia heterozygous for R496W and D374Y mutations were 12.8-fold (p=0.015) and 3.4-fold (p=0.03) higher than that in mutant and wild-type genotype, respectively. Additionally, in the entire study group (n=401), PCSK9 R496W and D374Y mutation carriers had increased total cholesterol (p=0.021), triglycerides (p=0.0001), HDL cholesterol (p=0.028), and low-density lipoproteins (LDL) cholesterol (p=0.028) levels. However, F216L (rs28942112) and S127R (rs28942111) mutations were not detected in patients with primary dyslipidemia and healthy controls. CONCLUSION: We conclude that the PCSK9 R496W (rs374603772) and D374Y (rs137852912) GOF mutations may be significant risk factors in the development of primary dyslipidemia and may have significant impact on lipid parameters in general population.
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Dislipidemias/genética , Predisposición Genética a la Enfermedad , Proproteína Convertasa 9/genética , Estudios de Casos y Controles , ADN/análisis , Dislipidemias/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mutación , Reacción en Cadena de la Polimerasa , Triglicéridos/sangre , Turquía , Población Blanca/genéticaRESUMEN
Resistance to chemotherapeutics including platinum agents, nucleoside analogues, topoisomerase and microtubule inhibitors is the most important problem in the treatment of many cancer. Several resistance mechanisms has been described for each group and researchers have been trying to develop new approaches to overcome chemoresistance. miRNAs are the regulators of gene transcription at the post-transcriptional level. Dysregulated miRNAs have been reported in cancer tumorigenesis, diagnosis and prognosis and they are promising new therapeutic approaches to improve diagnosis and predictions of outcomes and response to chemotherapy. In this article, we summarized the results from 2015 January to 2017 June which explore the roles of miRNAs in the development of the chemotherapy resistance.
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Resistencia a Antineoplásicos/genética , MicroARNs/metabolismo , Animales , Antineoplásicos/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Humanos , MicroARNs/genética , Modelos BiológicosRESUMEN
OBJECTIVE: The polymorphisms/mutations of genes encoding proteins and enzymes involved in lipoprotein metabolism play important roles in the development of diabetic dyslipidemia. The aim of our study was to investigate the effects of LPL (rs320), LIPC (rs2070895), SCARB1 (rs5888), LCAT (rs2292318), CETP (rs708272), ADIPOQ (rs1501299), RETN (rs3745367), PON1 (rs662), and MNSOD (rs4880) gene polymorphisms on lipid metabolism and diabetic dyslipidemia. METHODS: This case-control study included 217 patients with diabetic dyslipidemia and 212 healthy age- and gender-matched individuals. Genomic DNA isolation was performed from blood samples, and genotype analysis was performed using melting curve analysis on a LightCycler® 480 Instrument. The chi-square test was used to compare genotype distribution and allele frequencies between the groups. RESULTS: Significant associations were observed between LPL (rs320) (p<0.001), LIPC (rs2070895) (p<0.001), SCARB1 (rs5888) (p<0.001), LCAT (rs2292318) (p<0.001), CETP (rs708272) (p<0.001), ADIPOQ (rs1501299) (p=0.01), RETN (rs3745367) (p<0.001), and MNSOD (rs4880) (p<0.001) polymorphisms and diabetic dyslipidemia. However, no association was observed between PON1 (rs662) polymorphisms and diabetic dyslipidemia (p=0.611). CONCLUSION: LPL (rs320), LIPC (rs2070895), SCARB1 (rs5888), LCAT (rs2292318), CETP (rs708272), ADIPOQ (rs1501299), RETN (rs3745367), and MNSOD (rs4880) polymorphisms play an important role in basic molecular metabolism in diabetic dyslipidemia. Therefore, these polymorphisms may be used as a predictive marker for diabetic dyslipidemia in high-risk patients.
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Diabetes Mellitus Tipo 2 , Dislipidemias/genética , Predisposición Genética a la Enfermedad , Metabolismo de los Lípidos/genética , Lípidos/sangre , Polimorfismo Genético , Biomarcadores , Estudios de Casos y Controles , Dislipidemias/sangre , Femenino , Humanos , Lipoproteína Lipasa/sangre , Lipoproteína Lipasa/genética , Masculino , Persona de Mediana Edad , Turquía , Población BlancaRESUMEN
BACKGROUND: Flavopiridol a semisynthetic flavone that inhibits cyclin-dependent kinases (CDKs) and has growth-inhibitory activity and induces a blockade of cell-cycle progression at G1-phase and apoptosis in numerous human tumor cell lines and is currently under investigation in phase II clinical trials. Cancer stem cells (CSCs) are comprised of subpopulation of cells in tumors that have been proposed to be responsible for recurrence and resistance to chemotherapy. The aim of the present study was to investigate the effects of flavopiridol in cancer stem cell cytoskeleton, cell adhesion, and epithelial to mesenchymal transition in CSCs. METHODS: The cells were treated with flavopiridol to determine the inhibitory effect. Cell viability and proliferation were determined by using the WST-1 assay. Caspase activity and immunofluorescence analyses were performed for the evaluation of apoptosis, cell cytoskeleton, and epithelial-mesenchymal transition (EMT) markers. The effects of flavopiridol on the cell cycle were also evaluated. Flow cytometric analysis was used to detect the percentages of CSCs subpopulation. We analyzed the gene expression patterns to predict cell cycle and cell cytoskeleton in CSCs by RT-PCR. RESULTS: Flavopiridol-induced cytotoxicity and apoptosis at the IC50 dose, resulting in a significant increase expression of caspases activity. Cell cycle analyses revealed that flavopiridol induces G1 phase cell cycle arrest. Flavopiridol significantly decreased the mRNA expressions of the genes that regulate the cell cytoskeleton and cell cycle components and cell motility in CSCs. CONCLUSION: Our results suggest that Flavopiridol has activity against lung CSCs and may be effective chemotherapeutic molecule for lung cancer treatment.
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Antígeno AC133/metabolismo , Carcinoma de Células Escamosas/patología , Flavonoides/farmacología , Receptores de Hialuranos/metabolismo , Neoplasias Pulmonares/patología , Células Madre Neoplásicas/efectos de los fármacos , Piperidinas/farmacología , Antígeno AC133/efectos de los fármacos , Antineoplásicos/farmacología , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/metabolismo , Caspasas/biosíntesis , Caspasas/genética , Línea Celular Tumoral , Proliferación Celular , ADN de Neoplasias/genética , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Receptores de Hialuranos/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/metabolismo , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Reacción en Cadena de la PolimerasaRESUMEN
Similarities and differences in the cell cycle components, apoptosis and cytoskeleton-related molecules among mouse skin fibroblast cells (MSFs), mouse squamous cell lung carcinomas (SqCLCs) and mouse embryonic stem cells (mESCs) are important determinants of the behaviour and differentiation capacity of these cells. To reveal apoptotic pathways and to examine the distribution and the role of cell cycle-cell skeleton comparatively would necessitate tumour biology and stem cell biology to be assessed together in terms of oncogenesis and embryogenesis. The primary objectives of this study are to investigate the effects of flavopiridol, a cell cycle inhibitor, and geldanamycin, a heat shock protein inhibitor on mouse somatic, tumour and embryonic stem cells, by specifically focusing on alterations in cytoskeletal proteins, cell polarity and motility as well as cell cycle regulators. To meet these objectives, expression of several genes, cell cycle analysis and immunofluorescence staining of intracellular cytoskeletal molecules were performed in untreated and flavopiridol- or geldanamycin-treated cell lines. Cytotoxicity assays showed that SqCLCs are more sensitive to flavopiridol than MSFs and mESCs. Keratin-9 and keratin-2 expressions increased dramatically whereas cell cycle regulatory genes decreased significantly in the flavopiridol-treated MSFs. Flavopiridol-treated SqCLCs displayed a slight increase in several cell cytoskeleton regulatory genes as well as cell cycle regulatory genes. However, gene expression profiles of mESCs were not affected after flavopiridol treatment except the Cdc2a. Cytotoxic concentrations of geldanamycin were close to each other for all cell lines. Cdkn1a was the most increased gene in the geldanamycin-treated MSFs. However, expression levels of cell cytoskeleton-associated genes were increased dramatically in the geldanamycin-treated SqCLCs. Our results revealing differences in molecular mechanisms between embryogenesis and carcinogenesis may prove crucial in developing novel therapeutics that specifically target cancer cells.
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Apoptosis/efectos de los fármacos , Benzoquinonas/farmacología , Células Madre Embrionarias/efectos de los fármacos , Flavonoides/farmacología , Lactamas Macrocíclicas/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Piperidinas/farmacología , Actinas/análisis , Animales , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Transición Epitelial-Mesenquimal , Fibroblastos/efectos de los fármacos , Flavonoides/uso terapéutico , Queratina-2/análisis , Neoplasias Pulmonares/patología , Ratones , Piperidinas/uso terapéuticoRESUMEN
Platinum-based chemotherapies have long been used as a standard treatment in non-small cell lung cancer. However, cisplatin resistance is a major problem that restricts the use of cisplatin. Deregulated cell death mechanisms including apoptosis and autophagy could be responsible for the development of cisplatin resistance and miRNAs are the key regulators of these mechanisms. We aimed to analyse the effects of selected miRNAs in the development of cisplatin resistance and found that hsa-miR-15a-3p was one of the most significantly downregulated miRNAs conferring resistance to cisplatin in Calu1 epidermoid lung carcinoma cells. Only hsa-miR-15a-3p mimic transfection did not affect cell proliferation or cell death, though decreased cell viability was found when combined with cisplatin. We found that induced expression of hsa-miR-15a-3p via mimic transfection sensitised cisplatin-resistant cells to apoptosis and autophagy. Our results demonstrated that the apoptosis- and autophagy-inducing effects of hsa-miR-15a-3p might be due to suppression of BCL2, which exhibits a major connection with cell death mechanisms. This study provides new insights into the mechanism of cisplatin resistance due to silencing of the tumour suppressor hsa-miR-15a-3p and its possible contribution to apoptosis, autophagy and cisplatin resistance, which are the devil's triangle in determining cancer cell fate.
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Antineoplásicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/genética , Cisplatino/farmacología , Resistencia a Antineoplásicos/genética , Neoplasias Pulmonares/genética , MicroARNs/genética , Apoptosis/efectos de los fármacos , Apoptosis/genética , Western Blotting , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Técnica del Anticuerpo Fluorescente , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa , TransfecciónRESUMEN
A new platform for drug, gene and peptide-protein delivery is emerging, under the common name of "extracellular vesicles". Extracellular vesicles (EVs) are 30-1000 nm-sized cell-derived, liposome-like vesicles. Current research on EVs as nano-delivery systems for small-molecule drugs and genetic material, reveal that these tiny, biologically-derived vesicles carry a great potential to boost the efficacy of many therapeutic protocols. Several features of EVs; from efficacy to safety, from passive to active targeting ability, the opportunity to be biologically or chemically labelled, and most importantly, their eobiotic origin make them promising candidate for development of the next generation personalized nanomedicines. The aim of this article is to provide a view on the current research in which EVs are used as drug/genetic material delivery systems. Their application areas, drug loading and targeting strategies, and biodistribution properties are discussed.
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Sistemas de Liberación de Medicamentos/tendencias , Vesículas Extracelulares/metabolismo , Técnicas de Transferencia de Gen/tendencias , Nanomedicina/tendencias , Nanopartículas/metabolismo , Animales , Membrana Celular/efectos de los fármacos , Membrana Celular/genética , Membrana Celular/metabolismo , Micropartículas Derivadas de Células/genética , Micropartículas Derivadas de Células/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Exosomas/genética , Exosomas/metabolismo , Vesículas Extracelulares/genética , Humanos , Nanomedicina/métodos , Nanopartículas/administración & dosificación , Distribución Tisular/efectos de los fármacos , Distribución Tisular/fisiologíaRESUMEN
AIM: Folate metabolism is fundamental to several biological functions and required for cell replication, division, and survival. The mammalian folic acid cycle is highly complex and the enzymes, methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR), and methionine synthase reductase (MTRR), have crucial roles in this metabolic pathway. The common polymorphisms of the MTHFR (C677T and A1298C), MTRR (A66G), and MTR (A2756G) enzymes are well documented as folate deficiency-related disorders, but their roles have not been examined in acromegalic patients. The aim of this study was to compare the genotypic distribution of these gene polymorphisms between patients with acromegaly and controls and explore whether these polymorphisms were associated with biochemical and hormonal parameters in acromegaly. We examined 91 acromegaly patients and 112 healthy subjects who were compared in terms of age and gender. Blood specimens of the subjects were collected in tubes containing ethylenediaminetetraacetic acid. Genomic DNA was isolated from peripheral blood leukocytes and genotyping of the MTHFR (C677T and A1298C) gene polymorphisms was assessed by melting temperature analyses after real-time polymerase chain reaction (PCR), whereas MTRR A66G and MTR A2756G gene polymorphism analyses were performed by PCR/restriction fragment length polymorphism from the isolated DNA of the subjects. RESULTS: MTHFR-677TT genotype frequency was significantly higher in the acromegaly group than the control group (p=0.017), and a significant increase was found in fibrinogen (p=0.032) levels in 677TT-carrying acromegaly patients. MTRR-66AA genotype was significantly higher in the control group than the acromegaly group (p=0.004). Total cholesterol (p=0.048) and C-reactive protein (p=0.046) levels decreased significantly in 66AA genotypes. Although MTR-2756AG genotype frequency was not different between the control and acromegaly groups, 2756AG genotype-carrying individuals have higher left carotid intima-media thickness levels within the patient group. CONCLUSION: Our results suggest that polymorphisms of the genes encoding the folic acid metabolism enzymes affect biochemical parameters in acromegaly and this may result in predispositions to some complications associated with folate metabolism and acromegaly.
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5-Metiltetrahidrofolato-Homocisteína S-Metiltransferasa/genética , Acromegalia/genética , Ferredoxina-NADP Reductasa/genética , Ácido Fólico/metabolismo , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Acromegalia/metabolismo , Acromegalia/patología , Adulto , Grosor Intima-Media Carotídeo , Estudios de Casos y Controles , Femenino , Ácido Fólico/genética , Estudios de Asociación Genética , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo Genético , Reacción en Cadena en Tiempo Real de la Polimerasa , Factores de RiesgoRESUMEN
PURPOSE: Mesenchymal stem cells (MSCs) represent a new approach to the treatment of several neoplastic or non-neoplastic disorders. Their potential to repair damaged tissues through trans differentiation in conjunction with their immunomodulatory ability made them promising candidates for cell-based immunotherapy and regenerative medicine. In the present study, we aimed to determine the effects of MSCs on proliferation, apoptosis and gene expression profile of the acute lymphoblastic leukemia (ALL) cell line CCRF-CEM. METHODS: The experiments were performed after MSCs and CCRF-CEM cells were co-cultured for 72 hrs. We analyzed the gene expression patterns to predict oncogenic pathway dysregulation in the cell groups by quantitative RT-PCR and immunohistochemical staining. RESULTS: Cell proliferation was significantly inhibited in co-cultured CCRF-CEM cells compared to the control. Furthermore, growth factors, p53, Bax and Caspase-9 expressions were increased and cell-signaling gene expressions decreased significantly. Despite increased levels of growth factors (CTGF, VEGF, FGF, EGFR), the increased apoptosis level was triggered by p53/ Bax. CONCLUSION: In this study we have shown that human MSCs have inhibitory effect on their neighboring malignant leukemia cells.
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Proliferación Celular , Células Madre Mesenquimatosas , Leucemia-Linfoma Linfoblástico de Células Precursoras , Apoptosis , Línea Celular Tumoral , HumanosRESUMEN
PURPOSE: Increased activation of the JAK-STAT signaling pathway is frequently observed in several primary cancers as well as cancer cell lines. Thus, targeting JAK-STAT pathway components by different molecular-biologic approaches in the search for new anticancer therapies has become widespread and resulted in encouraging outcomes. In this study, the effects of chemically modified anti-STAT3 small interfering (si)RNAs on cell viability, proliferation and apoptosis of parental and cisplatin resistant non-small cell lung cancer (NSCLC) cells were investigated with the aim to provide a new therapeutic strategy for overcoming cisplatin resistance in lung cancer. METHODS: The parental NSCLC cell line Calu1 and its cisplatin- resistant subline CR-Calu1 were used to study the effects of STAT3 suppression with chemically modified anti-STAT3 siRNAs. STAT3 gene and protein expressions were analyzed by real-time (RT) quantitative (q) PCR and Western blot, respectively. Apoptosis was evaluated by Caspase-3 activity and cell death assays. RESULTS: STAT3 messenger (m)RNA and protein expression were significantly increased in CR-Calu1 cells and suppressing its expression with specific siRNAs increased the rate of apoptosis through Caspase-3 activation. STAT3 suppression also significantly increased cisplatin sensitivity of Calu1 and CR-Calu1 cells after transfection with STAT3 siRNAs. CONCLUSIONS: NSCLC cells could be sensitized to cisplatin by targeting STAT3 with chemically modified siRNAs together, a fact which was accompanied with increased apoptosis.
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Carcinoma de Pulmón de Células no Pequeñas/genética , Resistencia a Antineoplásicos/genética , ARN Interferente Pequeño/genética , Factor de Transcripción STAT3/genética , Apoptosis , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular , Cisplatino/administración & dosificación , Humanos , ARN Interferente Pequeño/química , Factor de Transcripción STAT3/metabolismoRESUMEN
PURPOSE: Capsaicin, an ingredient of red chili pepper, has possible tumorigenicity/genotoxicity properties. We aimed to determine the effects of capsaicin on the proliferation and gene expression profiles of acute lymphoblastic leukemia (ALL) CCRF-CEM cell line. METHODS: Cell viability and IC50 dose was determined by WST cytotoxicity assay. qRT-PCR, immunohistochemical staining and western blot methods were used to determine target genes' expression levels. Apoptosis was evaluated by measuring the caspase-3 activity. RESULTS: Capsaicin inhibited the proliferation of CCRFCEM cells in a dose-dependent manner. Increased mRNA expressions of caspase gene family members, activated caspase-3 and decreased mRNA and protein expression of BCL-2 gene indicated apoptotic response to capsaicin. Moreover capsaicin treatment suppressed significantly the expression of the key cell signaling pathways of KRAS, AKT, GAB2, PTPN11, BRAF, INPP5D, MAPK7. CONCLUSION: Capsaicin induces apoptosis in CCRF-CEM cells and this response is associated with downregulation of cell signaling pathways.
Asunto(s)
Apoptosis/efectos de los fármacos , Capsaicina/farmacología , Proliferación Celular/efectos de los fármacos , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Caspasa 3/biosíntesis , Caspasa 3/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Proteínas Proto-Oncogénicas c-bcl-2/genética , ARN Mensajero/biosíntesis , Transducción de Señal/efectos de los fármacosRESUMEN
OBJECTIVE: Hypertrophic cardiomyopathy (HCM) is a disease of the myocardium with an autosomal-dominant pattern of inheritance mainly caused by single heterozygous mutations in sarcomere genes. In this study we aimed to detect the presence of R403QLW, V606M, K615N, and R663H mutations in beta-myosin heavy-chain gene (MYH7) and figure out the genotype-phenotype correlations in Turkish patients with HCM. METHODS: This case-control study based on genotype-phenotype correlation included 69 patients (mean age, years: 50±13.16) diagnosed with HCM constituting the study group and 50 healthy individuals (mean age, years: 52±1.4) constituting the control group. DNA was extracted from peripheral blood and the genotyping of mutations was performed by real-time PCR technique and high resolution melting analysis. Associations between categoric variables were determined using chi-square tests. Differences between two groups were compared with unpaired Student's t-test for continuous variables. RESULTS: None of the patients in the HCM group were carrying the index mutations. One healthy individual was found to be heterozygous for the R663H mutation with mildly abnormal IVS and LVPW thickness. The allele frequency for R663H (G>A) mutation was found to be 0.01% in control group. CONCLUSION: We performed a mutational screening of 6 HCM-associated mutations in 69 Turkish HCM patients (not previously studied except R403Q). There was no significant difference in the prevalence of the mutations between the patients with HCM and the healthy controls (p>0.05).
Asunto(s)
Cardiomiopatía Hipertrófica/genética , Cadenas Pesadas de Miosina/genética , Cardiomiopatía Hipertrófica/sangre , Cardiomiopatía Hipertrófica/diagnóstico por imagen , Estudios de Casos y Controles , Análisis Mutacional de ADN , Cartilla de ADN , Ecocardiografía , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Turquía , Población BlancaRESUMEN
Cardiovascular diseases (CVDs) remain the main cause of morbidity and mortality around the world. A common polymorphism c.677C>T has been identified in the gene coding for methylenetetrahydrofolate reductase (MTHFR), which is involved in the remethylation of homocysteine, and may predispose to CVDs. A meta-analysis was performed to estimate the risk of CVDs associated with MTHFR c.677C>T in Turkish population. Published studies were retrieved from PubMed, Science Citation Index/Expanded, Google Scholar, Turkish Medline, and the Turkish Council of Higher Education Theses Database. For each study, we calculated odds ratios and 95 % confidence intervals (CI), assuming frequency of allele and homozygote comparison, dominant and recessive genetic models. Thirty-one separate studies were included and 2.780 cases/3.022 controls were involved in the current meta-analysis. Significant association was found between c.677C>T polymorphism and risk of CVD when all studies pooled with random-effects model for T versus C (OR 1.33; 95 % CI 1.11-1.59; p = 0.002), TT vs. CC (OR 1.87; 95 % CI 1.35-2.60; p = 3.53E-04), TT+CT vs. CC (OR 1.32; 95 % CI 1.06-1.64; p = 0.014) and TT vs. CT+CC (OR 1.75; 95 % CI 1.29-2.37; p = 6.57E-04). Further analysis indicated the significant association between methylenetetrahydrofolate reductase (MTHFR) TT genotype and groups with venous thrombosis, peripheral arterial thrombosis, acute MI/MI. No publication bias was observed in any comparison model. Our results of meta-analysis suggest that MTHFR c.677C>T polymorphism is associated with the CVDs in Turkish population.
Asunto(s)
Enfermedades Cardiovasculares/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Enfermedades Cardiovasculares/enzimología , Estudios de Casos y Controles , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Humanos , Oportunidad Relativa , Polimorfismo de Nucleótido Simple , Factores de Riesgo , TurquíaRESUMEN
The aim of this study was to investigate the effects of experimentally induced diabetes on (a) germ cells, (b) in vitro fertilization (IVF) success rate, and (c) gap junction and cell adhesion molecule gene and protein expressions during the early blastocyst period. Germ cells were obtained from healthy and diabetic rats, analyzed for number, motility, and morphology, and used for IVF. After reaching the early blastocyst stage, the expressions of genes encoding gap junction proteins and cell adhesion molecules were analyzed by quantitative RT-PCR. Histomorphologically and immunohistochemically analyses were also performed. Diabetes significantly affected sperm number and motility and the development of oocytes. Gene expressions of ß -catenin and connexin family members and protein expressions of E-cadherin and connexin-43 significantly decreased in groups including germ cells isolated from diabetic rats. Connective tissue growth factor expression increased in groups that included sperm cells isolated from diabetic male rats, whereas mucin-1 expression increased in the group that included oocytes isolated from diabetic female rats paired with sperm cells isolated from healthy male rats. In summary, experimentally induced diabetes was found to influence gap junctions, cell adhesion molecules, and associated proteins which all have important roles in germ cell maturation, fertilization, and development.
RESUMEN
The cytotoxicity of nine soft and hard lining materials (Mollosil Plus, Ufi Gel SC, Visco-gel, Molloplast-B, GC Tissue Conditioner, Vertex Rapid Simplified, GC Reline Hard, Vertex Self-Curing, Ufi Gel hard C) was evaluated using human gingival fibroblasts (HGFs). Twelve disk samples per lining material were prepared and incubated for 24, 48, 72, and 96 h. Cytotoxicity of each lining material's extract on cultured HGFs was measured using XTT assay. Data were analyzed using one-way ANOVA, post hoc Dunnett's T3 and Bonferroni tests at a significance level of p<0.05. At all incubation periods, all the hard lining materials (Vertex-SC, GC Reline Hard, Vertex-RS, and Ufi Gel hard C) showed cell viability higher than 90%. Among the soft lining materials, although there were no significant differences in cell viability among the different incubation periods for each lining material (p>0.05), autopolymerized acrylic-based GC Tissue Conditioner showed significantly lower cell viability than the other soft lining materials at each incubation period. Among the hard lining materials, there were no significant differences both among the materials and across all incubation periods for each lining material (p>0.05). In conclusion, all soft and hard liners exhibited good biocompatibility regardless of incubation time, except for GC Tissue Conditioner.
Asunto(s)
Alineadores Dentales/efectos adversos , Fibroblastos/efectos de los fármacos , Resinas Acrílicas/toxicidad , Análisis de Varianza , Materiales Biocompatibles/toxicidad , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Encía/citología , Humanos , Metilmetacrilatos/toxicidad , Elastómeros de Silicona/toxicidad , Estadísticas no ParamétricasRESUMEN
Acromegaly is a syndrome that results when the pituitary gland produces excess growth hormone after epiphyseal closure at puberty. Usually, subjects with acromegaly exhibit a 2- to 3-fold higher mortality rate from diseases that are associated with cardiovascular complications when compared to the normal population. In this study, we therefore aimed to evaluate whether a well-established cardiovascular risk factor, the Apolipoprotein E (Apo E) genotype, contributes to increased risk of cardiovascular complications in subjects with acromegaly. A total of 102 unrelated acromegaly subjects were prospectively included into this case-control association study and constituted our study group. The study group was comparable by age and gender with 200 unrelated healthy subjects constituting our control group. Genomic DNA was isolated from the peripheral blood leukocytes of all subjects and Apo E genotype (codon 112/158) was assessed by melting temperature analyses after using a real-time PCR protocol. The Apolipoprotein E4 allele was found at a significantly higher frequency in the study group when compared with the control group (P = 0.032). Subjects with the E2 allele, on the other hand, had significantly increased values in body mass index (P = 0.004), waist circumference (P = 0.001), C-reactive protein (CRP) (P < 0.001), and left-side carotid intima media thickness (P = 0.025). The Apolipoprotein E2 genotype might contribute to increased risk of cardiovascular complications in subjects with acromegaly since it is concurrently present with other cardiovascular risk factors such as the left-side carotid intima media thickness and CRP.