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Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignant cancer type worldwide. Although the therapeutic modalities currently used for patients with HNSCC improved in recent decades, HNSCC prognosis is still poor. Therefore, it is an urgent necessity to understand the pathogenesis of HNSCC, to develop novel and effective treatment strategies, and to characterize and identify the oncogenes that are responsible for an aggressive HNSCC phenotype. In this study, we aimed to better understand the roles of miR-1825 in the pathogenesis of HNSCC. We examined the impacts of miR-1825 deregulation on the cancer-associated phenotypes using in vitro tests evaluating cell viability, clonogenicity, cell migration, invasion, apoptosis, and stem cell characteristics. In addition, we investigated the effects of miR-1825 overexpression on the tumor formation capacity of head and neck cancer cells in vivo using nude mice. We searched for potential targets of miR-1825 using microarray analysis and luciferase assay. We found that miR-1825 expression is upregulated in head and neck cells and clinical tumor samples in comparison to corresponding controls, where it potentially acts as an oncogene. We, then, showed that ectopic miR-1825 overexpression promotes cellular phenotypes related to head and neck cancer progression in vitro and has a stimulating potential on cancer formation in vivo. We also identified FREM1 as a direct target of miR-1825 and demonstrated its reduced expression in HNSCC samples using immunohistochemistry analysis. Collectively, we suggest that the miR-1825/FREM1 axis serves as an important mediator of HNSCC development, where miR-1825 acts as an oncogene.
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Head and neck squamous cell carcinoma (HNSCC) is one of the most aggressive neoplasms, which requires more effective prevention and treatment modalities. Previous studies found that protein O-fucosyltransferase 1 (POFUT1) upregulation promotes carcinogenesis, although the potential roles, underlying molecular mechanisms, and biological implications of POFUT1 in HNSCC were not investigated. In this study, in silico analyses referred POFUT1 as a potential oncogene in HNSCC. Further analysis of tumor and normal tissue samples as well as HNSCC cells with quantitative real-time polymerase chain reaction, Western blot analysis, and immunohistochemistry showed significant overexpression of POFUT1 in HNSCC clinical tumor tissue specimens and cell lines compared to corresponding controls. In vitro investigations revealed that overexpression of POFUT1 promoted phenotypes associated with cancer aggressiveness and its knockdown in HNSCC cells suppressed those phenotypes. Further xenograft experiments demonstrated that POFUT1 is an oncogene in vivo for HNSCC. Immunohistochemical analysis with human clinical samples and cancer cell-dorsal root ganglion ex-vivo coculture model showed that deregulation of POFUT1 is involved in the perineural invasion of HNSCC cells. These results suggest POFUT1 expression as a potential prognostic marker for patients with head and neck cancer and highlight its potential as a target for HNSCC therapy, although more molecular clues are needed to better define the functions of POFUT1 related to HNSCC carcinogenesis.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Humanos , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Neoplasias de Cabeza y Cuello/genética , Fenotipo , Carcinogénesis , Línea Celular Tumoral , Proliferación Celular/genéticaRESUMEN
OBJECTIVES: Diets and nutritional habits are critical during carcinogenic processes, where a diet poor in fruits and vegetables and rich in meat and other foods of animal origin facilitates carcinogenesis. In this study, we aimed at investigating the possible involvement of vitamin D deficiency (VDD) and high cholesterol (HC) together in oral squamous cell carcinoma (OSCC) through modulating glycolysis. SUBJECTS AND METHODS: We compared total cholesterol, LDL, HDL, triglycerides, LDH, and vitamin D levels of OSCC patients and control individuals. We used GEO datasets for gene set enrichment and 4-nitroquinoline-1-oxide induced in vivo oral carcinogenesis models to investigate contribution of VDD and HC during carcinogenesis via possible modulation of glycolysis. RESULTS: We found that VDD and HC co-exist in OSCC patients, and deregulation of cholesterol and vitamin D levels results in enrichment of genes related to glycolysis. We, then, demonstrated that VDD and HC on their own and together facilitated the formation of larger tumors in 4NQO-induced in vivo cancer models, which are suppressed by glycolysis inhibition. CONCLUSION: We reported collaborative contribution of HC and VDD during oral carcinogenesis, which is mainly carried out via altering energy metabolism in tumor cells.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Boca , Deficiencia de Vitamina D , Ratas , Animales , Carcinoma de Células Escamosas/inducido químicamente , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Neoplasias de la Boca/genética , Carcinogénesis/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello , Vitamina D , Glucólisis , Deficiencia de Vitamina D/complicacionesRESUMEN
Peri-implantitis develops in 43.3% of implant patients, which affects tissues around the implant that may ultimately cause implant loss if not treated properly. Due to difficulties in detecting peri-implantitis in its early phases, implant failures are constantly on the rise. Therefore, new specific molecular markers need to be identified to prevent or limit disease progression in peri-implantitis patients. We investigated levels of CXCL9, CXCL12, and CXCL14 in saliva samples of 45 patients with commercially pure grade 4/5 Titanium-Aluminum-Vanadium implants. We analyzed the correlation of the chemokine levels using Pearson's Correlation test and investigated their power to discriminate peri-implantitis vs. non-peri-implantitis patients using receiver operating characteristic analysis. Our in silico investigation revealed CXCL9, CXCL12, and CXCL14 as predicted targets of miR-4484, which has been demonstrated as a powerful biomarker candidate for early detection of peri-implantitis in our previous study. We measured high CXCL9 and low CXCL14 levels in the saliva of peri-implantitis patients. We also reported that the CXCL14 level showed a significant positive correlation with miR-4484. Besides, CXCL14 together with miR-4484 in saliva differentiated peri-implantitis patients from non-peri-implantitis individuals with 100% success. We offer differential expressions of CXCL14 and miR-4484 in the saliva of patients with peri-implantitis as potential salivary biomarkers for early detection of this disease.
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Prostate cancer (PCa) is one of the most prevalent cancer types among males. Differential expression of microRNAs is associated with various cancers including PCa. Although mature microRNAs are preferentially located in the cytoplasm, several studies identified mature human microRNAs in purified nuclei and miR-145 has been found to be predominantly expressed in the nuclei of benign tissues compared to tumor lesions. However, the nuclear functions of miR-145 are yet limited. Here, we aimed at investigating the inductive role of miR-145 on the expression of Semaphorin 3A (SEMA3A) in PCa cell lines. To study the regulatory potential of miR-145 in the transcriptional level in PCa, we overexpressed miR-145 in PC3 and DU145 cells, and confirmed its upregulation by quantitative-real-time-PCR. Then we investigated the tumor suppressor potential of miR-145 upon inducing SEMA3A expression using cell viability assay, western blot analysis, Chromatin Immunoprecipitation assay and luciferase reporter assay. Our results revealed that p53, miR-145, and SEMA3A expressions are significantly downregulated in PC3 and DU145 cells compared to nontumorigenic prostate epithelial PNT1a cells. miR-145 overexpression in PCa cells induced the expression of SEMA3A at both messenger RNA and protein levels. Furthermore, increased miR-145 expression enriched RNA Pol-II antibody on the promoter of SEMA3A and induced luciferase activity controlled by SEMA3A promoter. In this study, we showed that the functions of miR-145 are not limited to gene silencing, and found that it may lead to changes in gene expression in the transcriptional level.
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MicroARNs/genética , Neoplasias de la Próstata/genética , Semaforina-3A/metabolismo , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Expresión Génica/genética , Perfilación de la Expresión Génica/métodos , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Masculino , MicroARNs/metabolismo , Regiones Promotoras Genéticas/genética , Próstata/metabolismo , Próstata/patología , Neoplasias de la Próstata/metabolismo , Semaforina-3A/genética , Transcriptoma/genéticaRESUMEN
Laryngeal squamous cell carcinoma (LSCC) is the second most common malignancy of the head and neck region in the USA with a declining 5-year survival rate. Paclitaxel resistance of tumors including LSCC still stands as a vital cause for poor clinical outcome in patients. In the current study, our aim was to explore the expressions of ATP-binding cassette transporters and stemness associated genes in human epithelial type 2 (Hep-2) cells with paclitaxel resistance. Resistant cells were developed via treatment with increasing doses of paclitaxel to acquire four sub-lines resistant to one-, two-, four-, and eightfold concentrations of paclitaxel (1×, 2×, 4×, 8×). Then, we profiled the expressions of ten selected ABC transporters (ABCA5, ABCB1, ABCB6, ABCC1, ABCC2, ABCC3, ABCC5, ABCC10, ABCF2, and ABCG2) and four stem cell markers (SOX2, OCT4, KLF, and CXCR4) using quantitative real time polymerase chain reaction in paclitaxel resistant cells to look for a link between these markers and chemoresistance. We demonstrated that ABCB1 and ABCG2 expressions gradually elevated and reached a maximum level in Taxol 8× cells. Considering stem cell markers, KLF4 expression elevated significantly, as soon as parental cells acquired resistance to the lowest dose of paclitaxel and its expression elevated stepwise. Expression levels of other tested ATP-binding cassette transporters and stem cell markers also elevated, although at different steps of paclitaxel resistance acquisition. Our findings suggest that higher expressions of ABCB1, ABCG2, and KLF4 might be considered as putative indicators for paclitaxel resistance in LSCC patients.
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Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2/genética , Factores de Transcripción de Tipo Kruppel/genética , Proteínas de Neoplasias/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/tratamiento farmacológico , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Antineoplásicos/farmacología , Línea Celular Tumoral , Resistencia a Múltiples Medicamentos/genética , Resistencia a Antineoplásicos/genética , Células Epiteliales/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Factor 4 Similar a Kruppel , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Paclitaxel/efectos adversos , Paclitaxel/farmacología , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/patologíaRESUMEN
BACKGROUND: Although there are no sufficient data on association between oxidative stress and erectile dysfunction (ED), numerous studies have reported that imbalance between the formation of reactive oxygen species and body's antioxidant defenses may play a role in the pathogenesis of ED. AIM: The aim of this study was to determine and compare the oxidant and antioxidant status in patients with ED and healthy controls with a novel automated assay for thiol/disulphide homeostasis test. METHODS: Our study included 123 patients with ED and 90 healthy individuals. ED was evaluated by asking questions 1-5 and 15 of the International Index of Erectile Function form. In this study, we used Erel and Neselioglu's thiol/disulfide homeostasis test, which is one of the novel methods that can measure both variables of the oxidative/antioxidative balance individually and collectively. OUTCOMES: This method measured serum antioxidant (total thiol [toSH], native thiol [SH]) and oxidant (disulfide [SS]) levels. The statistical comparisons were performed between patients with ED (ED+ group) and without ED (ED- group) first and then within the ED+ group. After toSH, SH, and SS levels were determined; SS/toSH%, SS/SH%, and SH/toSH% levels were analyzed separately and compared statistically. RESULTS: We found a significant difference between ED- and ED+ groups in terms of toSH, SH, SS/toSH%, and SS/SH% ratios. SS parameters were increased in patients with ED, but there was no significant difference in terms of SS and SH/toSH% values. CLINICAL IMPLICATIONS: Clarification of the factors involved in the etiology of ED such as oxidative/antioxidative balance may open new grounds in the early diagnosis and treatment of the disease. STRENGTHS & LIMITATIONS: It is a prospective, randomized clinical study with the use of a novel, reliable, and fully automated technique. The limitations of the study are use of a subjective tool such as the International Index of Erectile Function, obtaining blood samples from the peripheral vein instead of penile cavernosal tissue, and relatively small sample size. CONCLUSION: The results of this study showed that thiol/disulfide homeostasis is altered in ED, and this imbalance may be a factor in its pathophysiology. We determined that as ED gets more severe, toSH and SH parameters decrease, whereas SS parameter increases. Micoogullari U, Karatas OF, Kisa E, et al. Thiol/Disulfide Homeostasis in Patients With Erectile Dysfunction. J Sex Med 2020;17:1934-1941.
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Disfunción Eréctil , Estrés Oxidativo , Disulfuros/metabolismo , Disfunción Eréctil/metabolismo , Homeostasis , Humanos , Masculino , Estudios Prospectivos , Compuestos de SulfhidriloRESUMEN
Prostate cancer (PCa) is one of the most common types of cancer in men. In several recent studies, chromosomal deletions in the q arm of chromosome 2, where ING5 resides within, have been identified in various cancer types including PCa. In this study, we investigate the role of ING5 as a tumor suppressor in PCa. We examined the expression level of ING5 in tissue samples and cell lines using quantitative real-time polymerase chain reaction and western blot analysis. We tested the in vitro tumor suppressor potential of ING5 in PC3 and LNCaP cells stably overexpressing it using cell viability, colony formation, migration, invasion, and apoptosis assays. We then investigated the effects of ING5 on the Akt and p53 signaling using western blot analysis. We show that ING5 is significantly downregulated in PCa tumor tissue samples and cell lines compared with the corresponding controls. In vitro assays demonstrate that ING5 effectively suppresses proliferative, clonogenic, migratory, and invasive potential and induce apoptosis in PCa cells. ING5 may potentially exert its anti-tumor potential by inhibiting AKT and inducing p53 signaling pathways. Our findings demonstrate that ING5 possesses tumor suppressor roles in vitro, pointing its importance during the prostatic carcinogenesis processes.
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The present study was carried out in the attempt to synthesize a new class of potential anticancer agents comprising eleven compounds (24-34) sharing the 3,5-diarylisoxazole as a core. The chemical structure of the new synthesized compounds was established by IR, 1H NMR, 13C NMR and elemental analysis. Their biological potential towards prostate cancer was evaluated by using cancer PC3 cells and non-tumorigenic PNT1a cells. Interestingly, compound 26 distinguished from others with a quite high selectivity value that is comparable to 5-FU. The binding mode of 26 towards Ribosomal protein S6 kinase beta-1 (S6K1) was investigated at a molecular level of detail by employing docking simulations based on GLIDE standard precision as well as MM-GBSA calculations.
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Antineoplásicos/farmacología , Isoxazoles/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/metabolismo , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Isoxazoles/síntesis química , Isoxazoles/metabolismo , Simulación del Acoplamiento Molecular , Células PC-3 , Unión Proteica , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismoRESUMEN
Resistance of laryngeal squamous cell carcinoma cells to traditional therapeutic regimens still remains to be a major reason for therapeutic failure in patients. In this study, we aimed at investigating the expression profiles of ATP-binding cassette (ABC) transporters and stem cell markers in 5-fluorouracil (5-FU) resistant laryngeal Hep-2 cells. We treated parental Hep-2 cells, with stepwise increased doses of 5-FU for almost 1 year to develop 5-FU resistant sub-lines with resistance against varying levels of 5-FU concentrations (4 sub-lines resistant to 1, 2, 4, and eightfold of 5-FU). Then, we measured the expression levels of 10 genes from ABC transporters family and 4 stem cell associated markers using quantitative reverse transcription polymerase chain reaction (qRT-PCR) to find out a potential relationship between these markers and chemoresistance. We found that stemness-associated markers had elevated expressions from the beginning of 5-FU resistance acquisition. Their expressions elevated stepwise while parental Hep-2 cells got resistance to higher doses of 5-FU. Expressions of tested ABC transporters (ABCA5, ABCB1, ABCB6, ABCC1, ABCC2, ABCC3, ABCC5, ABCC10 and ABCF2, and ABCG2) were also deregulated in 5-FU resistant Hep-2 cells. Although their expressions remained unaltered at the beginning of acquisition of resistance, expressions of ABC transporters except from ABCB6 increased significantly when cells became resistant to higher doses of 5-FU. Our results suggest that enrichment of cells with stemness characteristics and upregulation of ABC transporters might be amongst the crucial contributors of chemoresistance in laryngeal cancer cells.
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Transportadoras de Casetes de Unión a ATP/genética , Resistencia a Antineoplásicos/genética , Neoplasias Laríngeas/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Fluorouracilo/farmacología , Expresión Génica/genética , Regulación Neoplásica de la Expresión Génica/genética , Células Hep G2 , Humanos , Proteína 2 Asociada a Resistencia a Múltiples Medicamentos , Células Madre Neoplásicas/metabolismo , Células Madre/metabolismoRESUMEN
PURPOSE: Martsolf (MS) and Warburg micro syndromes (WARBM) are rare autosomal recessive inherited allelic disorders, which share similar clinical features including microcephaly, intellectual disability, brain malformations, ocular abnormalities, and spasticity. Here, we revealed the functions of novel mutations in RAB3GAP1 in a Turkish female patient with MS and two siblings with WARBM. We also present a review of MS patients as well as all reported RAB3GAP1 pathogenic mutations in the literature. METHODS: We present a female with MS phenotype and two siblings with WARBM having more severe phenotypes. We utilized whole-exome sequencing to identify the molecular basis of these syndromes and confirmed suspected variants by Sanger sequencing. Quantitative (q) RT-PCR analysis was carried out to reveal the functions of novel splice site mutation detected in MS patient. RESULTS: We found a novel homozygous c.2607-1G>C splice site mutation in intron 22 of RAB3GAP1 in MS patient and a novel homozygous c.2187_2188delinsCT, p.(Met729_Lys730delinsIleTer) mutation in exon 19 of RAB3GAP1 in the WARBM patients. We showed exon skipping in MS patient by Sanger sequencing and gel electrophoresis. qRT-PCR analysis demonstrated the reduced expression of RAB3GAP1 in the patient with the c.2607-1G>C splice site mutation compared to a healthy control individual. CONCLUSION: Here, we have studied two novel RAB3GAP1 mutations in two different phenotypes; a MS associated novel splice site mutation, and a WARBM1 associated novel deletion-insertion mutation. Our findings suggest that this splice site mutation is responsible for milder phenotype and the deletion-insertion mutation presented here is associated with severe phenotype.
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Anomalías Múltiples/genética , Anomalías Múltiples/patología , Empalme Alternativo , Catarata/congénito , Córnea/anomalías , Hipogonadismo/genética , Hipogonadismo/patología , Discapacidad Intelectual/genética , Discapacidad Intelectual/patología , Microcefalia/genética , Microcefalia/patología , Mutación , Atrofia Óptica/genética , Atrofia Óptica/patología , Proteínas de Unión al GTP rab3/genética , Catarata/genética , Catarata/patología , Niño , Córnea/patología , Femenino , Homocigoto , Humanos , Mutación INDEL , Masculino , Linaje , Fenotipo , Hermanos , TurquíaRESUMEN
Cell lines established from tumors are the most commonly used models in cancer research, and their use in recent years has enabled a greater understanding of the biology of cancer and the means to develop effective treatment strategies. Astroblastomas are uncommon neuroepithelial tumors of glial origin, predominantly affecting young people, mainly teenagers and children, predominantly females. To date, only a single study has reported that astroblastomas contain a large number of neural stem-like cells, which had only a partial proliferation capacity and differentiation. Our objective was to establish an astroblastoma cell line to investigate the presence of astroblastic cells and cancer stem-like cells. The migratory and invasion abilities of the cells were quantified with invasion and migration assays and compared to a glioblastoma cell line. The presence of stem cells was detected with surface-marker analysis by using flow cytometry, and measuring the differentiation ability with a differentiation assay and the self-renewal capacity with a sphere-forming assay. These characteristics may determine whether this novel cell line is a model for astroblastomas that may have stem-cell characteristics. With this novel cell line, scientists can investigate the molecular pathways underlying astroblastomas and develop new therapeutic strategies for patients with these tumors.
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Técnicas de Cultivo de Célula/métodos , Neoplasias Neuroepiteliales/patología , Células Madre Neoplásicas/citología , Línea Celular Tumoral , Separación Celular/métodos , Humanos , Masculino , Células Madre Neoplásicas/fisiología , Esferoides Celulares/citología , Esferoides Celulares/fisiología , Células Tumorales Cultivadas , Adulto JovenRESUMEN
The cancer stem-like cells (CSLCs) are tumorigenic cells promoting initiation, progression, and spread of the tumor. Accumulating evidences suggested the presence of CSLCs in distinct tumors including laryngeal squamous cell carcinoma (LSCC). MicroRNAs have been proposed as significant regulators of carcinogenesis, and several of them have been demonstrated to have direct roles in survival of CSLCs. In this study, we aimed to explore the role of miR-145, which is downregulated in LSCC, on cancer stem cell potency of laryngeal cancer cells. We initially showed the downregulation of miR-145 expression in tumor tissue samples and in CD133-enriched CSLCs. Quantitative reverse-transcription PCR (qRT-PCR) analysis of miR-145-transfected Hep-2 cells demonstrated the inhibitory role of miR-145 on stem cell markers like SOX2, OCT4, KLF4, and ABCG2. We, then, investigated the stem cell features of miR-145-overexpressing Hep-2 cells by sphere formation assay, single-cell cloning assay, and aldehyde dehydrogenase (ALDH) assay, which all demonstrated the inhibition of stem cell potency upon miR-145 overexpression. Further qRT-PCR analysis demonstrated altered expression of epithelial to mesenchymal transition markers in miR-145-overexpressing Hep-2 cells. In conclusion, we demonstrated the regulatory role of miR-145 in stem cell characteristics of Hep-2 cells. Based on these results, we propose that miR-145 might carry crucial roles in LSCC tumorigenesis, prognosis, metastasis, chemoresistance, and recurrence through regulating stem cell properties of tumor cells.
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Carcinoma de Células Escamosas/metabolismo , Neoplasias Laríngeas/metabolismo , MicroARNs/fisiología , Células Madre Neoplásicas/metabolismo , Adulto , Anciano , Aldehído Deshidrogenasa/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , Femenino , Humanos , Factor 4 Similar a Kruppel , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/patología , Masculino , Persona de Mediana Edad , Esferoides Celulares/metabolismoRESUMEN
Autism spectrum disorder (ASD) is one of the lifelong existing disorders. Abnormal methylation status of gene promoters of oxytonergic system has been implicated as among the etiologic factors of ASDs. We, therefore, investigated the methylation frequency of oxytocin receptor gene (OXTR) promoter from peripheral blood samples of children with autistic features. Our sample includes 66 children in total (22-94 months); 27 children with ASDs according to the DSM-IV-TR and the Childhood Autism Rating Scale (CARS) and 39 children who do not have any autistic like symptoms as the healthy control group. We investigated the DNA methylation status of OXTR promoter by methylation specific enzymatic digestion of genomic DNA and polymerase chain reaction. A significant relationship has been found between ASDs and healthy controls for the reduction of methylation frequency of the regions MT1 and MT3 of OXTR. We could not find any association in the methylation frequency of MT2 and MT4 regions of OXTR. Although our findings indicate high frequency of OXTR promoter hypomethylation in ASDs, there is need for independent replication of the results for a bigger sample set. We expect that future studies with the inclusion of larger, more homogeneous samples will attempt to disentangle the causes of ASDs.
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Trastorno del Espectro Autista/genética , Metilación de ADN/genética , Regiones Promotoras Genéticas/genética , Receptores de Oxitocina/genética , Niño , Preescolar , Femenino , Humanos , Lactante , MasculinoRESUMEN
BACKGROUND: Emerging evidences proposed that microRNAs are associated with regulation of distinct physio-pathological processes including development of normal stem cells and carcinogenesis. In this study we aimed to investigate microRNA profile of cancer stem-like cells (CSLCs) isolated form freshly resected larynx cancer (LCa) tissue samples. METHODS: CD133 positive (CD133+) stem-like cells were isolated from freshly resected LCa tumor specimens. MicroRNA profile of 12 pair of CD133+ and CD133- cells was determined using microRNA microarray and differential expressions of selvected microRNAs were validated by quantitative real time PCR (qRT-PCR). RESULTS: MicroRNA profiling of CD133+ and CD133- LCa samples with microarray revealed that miR-26b, miR-203, miR-200c, and miR-363-3p were significantly downregulated and miR-1825 was upregulated in CD133+ larynx CSLCs. qRT-PCR analysis in a total of 25 CD133+/CD133- sample pairs confirmed the altered expressions of these five microRNAs. Expressions of miR-26b, miR-200c, and miR-203 were significantly correlated with miR-363-3p, miR-203, and miR-363-3p expressions, respectively. Furthermore, in silico analysis revealed that these microRNAs target both cancer and stem-cell associated signaling pathways. CONCLUSIONS: Our results showed that certain microRNAs in CD133+ cells could be used as cancer stem cell markers. Based on these results, we propose that this panel of microRNAs might carry crucial roles in LCa pathogenesis through regulating stem cell properties of tumor cells.
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Biomarcadores de Tumor/metabolismo , Neoplasias Laríngeas/metabolismo , MicroARNs/metabolismo , Células Madre Neoplásicas/metabolismo , Antígeno AC133/metabolismo , Biomarcadores de Tumor/genética , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Laríngeas/patología , Masculino , MicroARNs/genética , Persona de Mediana Edad , Interferencia de ARN , Células Tumorales CultivadasRESUMEN
We aimed to perform functional analysis of miR-145-5p in prostate cancer (PCa) cells and to identify targets of miR-145-5p for understanding its role in PCa pathogenesis. PC3, DU145, LNCaP PCa, and PNT1a nontumorigenic prostate cell lines were utilized for functional analysis of miR-145-5p. Its overexpression caused inhibition of proliferation through apoptosis and reduced migration in PCa cells. SOX2 expression was significantly decreased in both mRNA and protein level in miR-145-5p-overexpressed PCa cells. We proposed that miR-145-5p, being an important regulator of SOX2, carries a crucial role in PCa tumorigenesis.
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Proliferación Celular , Regulación Neoplásica de la Expresión Génica/fisiología , MicroARNs/biosíntesis , Neoplasias de la Próstata/patología , Factores de Transcripción SOXB1/biosíntesis , Apoptosis/fisiología , Western Blotting , Línea Celular Tumoral , Proliferación Celular/fisiología , Humanos , Masculino , Análisis de Secuencia por Matrices de Oligonucleótidos , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
OBJECTIVES: The aim of this study is to evaluate the relationship of miR-21, nitric oxide (NOx) and endothelial nitric oxide synthase (eNOS) with subclinical atherosclerosis in carotid arteries by measuring carotid intima media thickness (CIMT) in patients with hypertension and healthy controls. DESIGN AND METHODS: A total of 28 hypertensive and 28 healthy controls were enrolled. MiR-21 expression was analyzed by quantitative reverse transcription-PCR and NOx, and eNOS levels were measured by ELISA assay. CIMT was evaluated by ultrasonography and CIMT ≥ 0.8 mm was accepted as increased CIMT (iCIMT). RESULTS: C-reactive protein (CRP) level, plasma miR-21 expression level and CIMT were found to be significantly higher in the hypertension group when compared to the control group (p = 0.009, p = 0.002 and p < 0.001, respectively). NOx and eNOS levels were significantly lower in the hypertension group compared to the control group (p < 0.001, both). MiR-21 level was positively correlated with the clinical systolic blood pressure, clinical diastolic blood pressure, CRP and CIMT. MiR-21 was also negatively correlated with NOx and eNOS. Eighteen patients with hypertension had iCIMT. MiR-21 and CRP levels were significantly higher (p < 0.001 and p = 0.001), whereas NOx and eNOS levels were significantly lower in patients with iCIMT (p < 0.001, both). CONCLUSION: The decreased levels of NOx and eNOS found in this study indicate the co-existence of endothelial dysfunction and hypertension once more. In the absence of microalbuminuria, the increased miR-21 expression in patients with iCIMT made us conclude that this miRNA might be involved in the early stages of atherosclerotic process in hypertensive patients.
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Aterosclerosis/genética , Presión Sanguínea/fisiología , Regulación de la Expresión Génica , Hipertensión/etiología , MicroARNs/genética , Óxido Nítrico Sintasa de Tipo III/genética , ARN/genética , Adulto , Aterosclerosis/sangre , Aterosclerosis/complicaciones , Arteria Carótida Externa/diagnóstico por imagen , Arteria Carótida Externa/fisiopatología , Grosor Intima-Media Carotídeo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Hipertensión/sangre , Hipertensión/genética , Masculino , MicroARNs/biosíntesis , MicroARNs/sangre , Persona de Mediana Edad , Óxido Nítrico Sintasa de Tipo III/biosíntesis , Óxido Nítrico Sintasa de Tipo III/sangre , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ultrasonografía Doppler en ColorRESUMEN
BACKGROUND: Cancer is one of the most common causes of human deaths worldwide. Nanotechnology has the potential to facilitate the detection, diagnosis, and treatment of cancer cases. Successful delivery of nucleic acids into cancer cells with the use of nanoparticles would be a significant improvement for medical and cellular biology. The use of nanoparticle-based vehicles in clinical treatment is considerably important for treating genetic disorders. Gold nanoparticles (AuNPs) have been suggested as therapeutic delivery tools for cancer. Because microRNAs (miRNAs), which induce post-transcriptional gene silencing, are deregulated in cancer cells, they are also considered as strong candidates for cancer therapy applications. In prostate and breast cancer, miR-145, a well-known tumor suppressor miRNA, is strongly downregulated in tumor tissues compared to their corresponding normal tissues. METHODS: In the present study, we aimed to use engineered AuNPs as nanocarrier platforms to deliver miRNAs to prostate/breast cancer cells. 13-nm AuNPs were modified with thiolated RNAs and then the miR-145 was hybridized to the RNAs that were chemically attached to the AuNPs. RESULTS: The results obtained in the present study demonstrate the efficient delivery of miR-145 to prostate/breast cancer cells. We also show that delivery was more efficient when the AuNP-RNA-miRNA carrier complex was formed at an elevated temperature of 72 °C. CONCLUSIONS: In conclusion, we show that AuNPs help the effective in vitro delivery of miR-145 into cancer cells.
Asunto(s)
MicroARNs/genética , Transfección , Neoplasias de la Mama , Femenino , Terapia Genética , Oro , Humanos , Células MCF-7 , Masculino , Nanopartículas del Metal , Nanopartículas , Neoplasias de la PróstataRESUMEN
Neurons and glia differentiate from multipotent precursors called neural stem cells (NSCs), upon the activation of specific transcription factors. In vitro, it has been shown that NSCs display very plastic features; however, one of the major challenges is to understand the bases of lineage restriction and NSC plasticity in vivo, at the cellular level. We show here that overexpression of the Gcm transcription factor, which controls the glial versus neuronal fate choice, fully and efficiently converts Drosophila NSCs towards the glial fate via an intermediate state. Gcm acts in a dose-dependent and autonomous manner by concomitantly repressing the endogenous program and inducing the glial program in the NSC. Most NSCs divide several times to build the embryonic nervous system and eventually enter quiescence: strikingly, the gliogenic potential of Gcm decreases with time and quiescent NSCs are resistant to fate conversion. Together with the fact that Gcm is able to convert mutant NSCs that cannot divide, this indicates that plasticity depends on temporal cues rather than on the mitotic potential. Finally, NSC plasticity involves specific chromatin modifications. The endogenous glial cells, as well as those induced by Gcm overexpression display low levels of histone 3 lysine 9 acetylation (H3K9ac) and Drosophila CREB-binding protein (dCBP) Histone Acetyl-Transferase (HAT). Moreover, we show that dCBP targets the H3K9 residue and that high levels of dCBP HAT disrupt gliogenesis. Thus, glial differentiation needs low levels of histone acetylation, a feature shared by vertebrate glia, calling for an epigenetic pathway conserved in evolution.
Asunto(s)
Cromatina/metabolismo , Proteínas de Unión al ADN/fisiología , Proteínas de Drosophila/fisiología , Regulación del Desarrollo de la Expresión Génica , Histonas/química , Células-Madre Neurales/citología , Neuroglía/citología , Factores de Transcripción/fisiología , Acetilación , Animales , Diferenciación Celular , División Celular , Linaje de la Célula , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Epigénesis Genética , Histonas/metabolismo , Inmunohistoquímica/métodos , Hibridación in Situ , Mitosis , Factores de Transcripción/metabolismo , Vertebrados/metabolismoRESUMEN
An increased pretreatment neutrophil-lymphocyte ratio (NLR) is associated with poor prognosis in colorectal, gastric, and ovarian cancer; malignant mesothelioma; and renal cell carcinoma. The present study aims to define the predictive value of preoperative peripheral blood count NLR in non-muscle-invasive bladder cancer (NMIBC) and muscle-invasive disease (MIBC) patients. There were in total 291 patients, 241 males and 50 females. Out of these, 156 male and 36 female patients were in the NMIBC group and 85 male and 14 female patients in the MIBC group. In the NMIBC group, 172 patients had low-grade and 20 high-grade papillary urothelial carcinoma. The mean age of the patients in the NMIBC group was 64 ± 13, ranging from 27 to 97. The mean age of the patients in MIBC group was 70.5 ± 10, ranging from 27 to 95. A statistically significant relation between patient ages and tumor invasiveness was determined (p = 0.023, 95 % confidence interval (CI) 63.3-66.7). The mean tumor size of the NMIBC group was 2.1 ± 1.09 (cm) (range 0.5-8), and of MIBC group 3.6 ± 1.5 (cm) (range 0.8-9). There was a statistically significant relation between the tumor size and invasiveness (p = 0.002, 95 % CI 2.8-4.4). In the NIMBC group, 149 (77.6 %) of them have NLR ≤ 2.5 and 43 (22.4 %) have NLR > 2.5. Also, in MIBC, 67 (67.7 %) of them have NLR ≤ 2.5 and 32 (32.3 %) have NLR > 2.5. The mean NLR in the NMIBC group was 2.4 ± 0.1 (range 0.08-6.49, 95 % CI 1.52-2.71) and in the MIBC 2.9 ± 0.2 (range 0.08-16.72, 95 % CI 1.67-2.97). In terms of NLR, there was a statistically significant difference between the NMIBC and MIBC groups (p = 0.028). Platelet-lymphocyte ratio (PLR) of the two groups was also analyzed. The PLR of the NMIBC group was 12.8 ± 15.1 (range 3.38-19.1) and of the MIBC 13.6 ± 8.78 (range 0.18-63), yet there was not any statistically significant difference (p = 0.810, 95 % CI 11.4-14.8) (Table 1). The correlation tests revealed a positive correlation between the age (r = 0.144, p = 0.024), tumor size (r = 0.193, p = 0.02), and tumor invasiveness NLR (r = 0.138, p = 0.031). NLR can be used to determine tumor invasiveness as a cost-effective, common, and simple biomarker in bladder cancer (BC).