RESUMO
This study was designed to evaluate serum LC3-II, BCL-2, IL-1ß, TGF-ß1, and podocin levels in. type 2 diabetes (T2DM) patients with renal dysfunction. MATERIALS: 176 Turkish subjects were enrolled, of whom 26 were healthy, and 150 had T2DM. PATIENTS: were classified according to albumin urea ratio: 88 patients had macroalbuminuria, 20. patients had microalbuminuria, and 42 had normoalbuminuria. T2DM patients were also. classified into three groups according to proteinuria and eGFR stages. RESULTS: Increased serum LC3-II levels in patients with T2DM with increased urinary albumin. extraction and impaired renal functions. There was a strong relationship between serum. LC3-II levels and serum BCL-2, IL-1ß, TGF-ß1, and Podocin levels. The efficiency of LC3- II as a diagnostic biomarker in the differential diagnosis of DM patients with. macroproteinuria from DM patients with normoproteinuria was 75.4%. CONCLUSIONS: It was thought that increased serum LC3-II levels in T2DM patients with impaired renal. functions may cause renal podocyte damage. In these patients, serum LC3-II levels can be. evaluated as a new biomarker to follow the development of renal damage.
Assuntos
Diabetes Mellitus Tipo 2 , Humanos , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Masculino , Feminino , Pessoa de Meia-Idade , Fator de Crescimento Transformador beta1/sangue , Biomarcadores/sangue , Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/fisiopatologia , Idoso , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Albuminúria/sangue , Interleucina-1beta/sangue , Adulto , Rim/fisiopatologia , Taxa de Filtração Glomerular , Proteínas de Membrana/sangue , Proteínas Associadas aos Microtúbulos , Peptídeos e Proteínas de Sinalização IntracelularRESUMO
PURPOSE: Behçet's disease (BD) is an autoimmune chronic systemic inflammatory disease characterized by a versatile clinical spectrum. Growth arrest specific protein 6 (GAS6)/soluble AXL (sAXL) signaling pathway draws attention in the resolution of inflammation, and its deficiency is associated with chronic inflammatory, autoimmune diseases, as well as clearance of apoptotic cells by phagocytes - efferocytosis. In this study, it was aimed to investigate whether GAS6/sAXL, interleukin (IL)-10, nitric oxide (NO), and BCL-2 levels were associated with inflammation and efferocytosis contributes to the pathogenesis of BD. METHODS: A total of 37 Behçet patients with ocular involvement and 30 healthy control subjects were included in this study. GAS6, sAXL, IL-10, NO, and BCL-2 levels were quantified using enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Serum GAS6, sAXL, IL-10, NO, and BCL-2 levels were significantly lower in patients with BD compared to the controls (P < 0.005, P < 0.001, P < 0.001, P < 0.001, and P < 0.001, respectively). In correlation analysis, research parameters decreased in patients with BD was significantly correlated with each other: GAS6-IL-10 (r = 0.585, P < 0.001), GAS6-BCL-2 (r = 0.541, P < 0.001), sAXL-BCL-2 (r = 0.696, P < 0.001), IL-10-NO (r = 0.717, P < 0.001), IL-10-BCL-2 (r = 0.759, P < 0.001), and NO-BCL-2 (r = 0.541, P < 0.001). CONCLUSION: In conclusion, decreased serum BCL-2 level may be an indicator of increased apoptosis in these patients and decreased levels of GAS6/sAXL, IL-10, and NO may indicate insufficient clearance of apoptotic bodies released as a result of increased apoptosis in BD patients.
Assuntos
Síndrome de Behçet , Biomarcadores , Ensaio de Imunoadsorção Enzimática , Peptídeos e Proteínas de Sinalização Intercelular , Interleucina-10 , Óxido Nítrico , Proteínas Proto-Oncogênicas c-bcl-2 , Proteínas Proto-Oncogênicas , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Receptor Tirosina Quinase Axl , Síndrome de Behçet/sangue , Síndrome de Behçet/diagnóstico , Biomarcadores/sangue , Peptídeos e Proteínas de Sinalização Intercelular/sangue , Interleucina-10/sangue , Óxido Nítrico/sangue , Proteínas Proto-Oncogênicas/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Receptores Proteína Tirosina Quinases/sangueRESUMO
Honeybee products consist of many substances, which have long been known for their medicinal and health-promoting properties. This study set out to appraise the protective potential of Egyptian propolis (EP) and bee venom (BV) separately or combined against total body irradiation (TBI) induced oxidative injury in rats. Besides, we assessed the bioactive components in EP and BV using HPLC and UPLC/ ESI-MS analysis in the positive ion mode. The animals were subjected to a source of gamma ionizing radiation at a dose of 6 Gy. Propolis and BV were administered independently and in combination before 14 days of γ-irradiation. Liver and kidney functions were estimated besides, DNA damage index (8- OHdG) by ELISA. Antioxidants, including glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were detected. Gene expression technique investigated for BAX, BCL2, and in plasma also miR125b expression in serum of rats. Besides, the histopathological for the brain, liver, kidney, and heart were investigated. In addition, lipid peroxidation was investigated in plasma and in the previous organs. The present results provide opportunities to advance the use of bee products as promising medicinal sources.
Assuntos
Venenos de Abelha/farmacologia , Raios gama/efeitos adversos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Própole/farmacologia , Protetores contra Radiação , Animais , Antioxidantes/metabolismo , Venenos de Abelha/administração & dosagem , Venenos de Abelha/química , Dano ao DNA/efeitos dos fármacos , Técnicas In Vitro , Própole/administração & dosagem , Própole/química , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Proteína Supressora de Tumor p53/sangue , Proteína X Associada a bcl-2/sangueRESUMO
Acute myocardial infarction (AMI) is a severe cardiovascular disease. AMI associated with coronary artery disease (AMI-CAD) is a subtype of AMI, composed of AMI patients caused by CAD. This study aimed to evaluate the diagnostic value of miR-96-5p in AMI induced by coronary artery disease. Expression of miR-96-5p and BCL2L13 was evaluated by serum samples and cells utilizing Western blot and RT-qPCR assays. The diagnostic value of miR-96-5p in AMI-CAD was analyzed with a receiver operating characteristic (ROC) curves. The correlation between miR-96-5p and BCL2L13 was examined by Spearman's correlation analysis. The level of oxidative stress and apoptosis were estimated via relative commercial kit, flow cytometry apoptosis assay and TUNEL staining assay. Our study discovered that miR-96-5p was down-regulated while BCL2L13 was up-regulated in patients with AMI-CAD. miR-96-5p was a potential diagnostic parameter, which may help distinguish AMI-CAD patients from healthy controls. In vitro experiments, miR-96-5p expression was down regulated while BCL2L13 was up-regulated in hypoxic cardiomyocytes. After confirming the targeted link of miR-96-5p to BCL2L13 using luciferase reporter and RNA pull down assays, we discovered that miR-96-5p overexpression may restore oxidative stress and cell apoptosis induced by hypoxia treatment in H9c2 cells; meanwhile, co-transfection with BCL2L13 overexpressing plasmid might partly countervail the ameliorative effects of miR-96-5p on oxidative stress and apoptosis. Collectively, miR-96-5p may function as a potential diagnostic biomarker for AMI-CAD patients, and the up-regulation of miR-96-5p would ameliorate AMI-associated cardiomyocytes injury by targeting BCL2L13, hence contributing to the clinical treatment of AMI-CAD.
Assuntos
Doença da Artéria Coronariana , MicroRNAs/sangue , Infarto do Miocárdio , Idoso , Animais , Biomarcadores/sangue , Hipóxia Celular/genética , Linhagem Celular , Doença da Artéria Coronariana/complicações , Doença da Artéria Coronariana/metabolismo , Feminino , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Infarto do Miocárdio/complicações , Infarto do Miocárdio/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Proto-Oncogênicas c-bcl-2/genética , RatosRESUMO
BACKGROUND: In the present study we investigated the levels of proapoptotic caspase-9 and antiapoptotic Bcl-2 proteins in the sera of children and adolescents with idiopathic epilepsy and tried to relate the findings to the patients' clinical parameters. METHODS: This retrospective study consisted of 118 children and adolescents with idiopathic epilepsy, categorized according to type and number of seizures, duration of the disease and the control of seizures and 30 age- and sex-matched controls. The relapse of seizures was taken into consideration. RESULTS: Mean serum level between Bcl-2 and caspase-9 was significantly higher only in Bcl-2 patients, compared to controls (P≤0.0001) and (P=0.987) respectively. Significant difference in Bcl-2 level was found among the different types of focal seizures. Caspase-9 level was statistically different in patients with two or more seizures per month compared to those with one seizure per month (P=0.048). No correlation was found between Bcl-2 and caspase-9 levels and age, gender, seizure frequency, total number of seizures and the duration of epilepsy. No significant difference was found in patients with and without drug treatment. CONCLUSIONS: Bcl-2 displays an association with apoptosis and highlights the potential of being a surrogate biomarker for active seizures and epilepsy. There is a significant difference in Bcl-2 serum level among the different types of focal seizures. Proapoptotic caspase-9 cannot act as a marker of active seizures and epilepsy. Caspase-9 serum level is increased acutely in controlled cases after a single relapse.
Assuntos
Caspase 9/sangue , Epilepsia , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Convulsões , Adolescente , Criança , Epilepsia/sangue , Epilepsia/tratamento farmacológico , Humanos , Estudos Retrospectivos , Convulsões/sangueRESUMO
BACKGROUND: There are scarce and contradictory data existing about B-cell lymphoma 2 (Bcl2), one of the Bcl2 family of anti-apoptotic proteins, in traumatic brain injury (TBI) patients. Thus, the objective of this study was to analyze whether blood concentrations of Bcl2 are associated with mortality. METHODS: Patients with isolated and severe TBI, defined as <10 points of the Injury Severity Score (ISS) in non-cranial aspects and <9 points in Glasgow Coma Scale (GCS), were included. This was an observational and prospective study carried out in five Intensive Care Units. Serum Bcl2 concentrations on day 1 of TBI were determined. RESULTS: Serum Bcl2 concentrations were lower (p < 0.001) in surviving patients (n = 59) compared to non-survivors (n = 24). We found an association between serum Bcl2 levels and mortality controlling for age and GCS (OR = 1.149; 95% CI = 1.056-1.251; p = 0.001) and controlling for computer tomography findings (OR = 1.147; 95% CI = 1.056-1.246; p = 0.001). CONCLUSIONS: This study reports for the first time an association between serum Bcl2 levels and 30-day mortality in TBI patients.
Assuntos
Lesões Encefálicas Traumáticas , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Reguladoras de Apoptose , Lesões Encefálicas Traumáticas/diagnóstico , Lesões Encefálicas Traumáticas/mortalidade , Humanos , Estudos ProspectivosRESUMO
Background: There are no data on circulating concentrations of sFas (proapoptotic protein of extrinsic pathway) and Bcl2 (antiapoptotic protein of intrinsic pathway) in COVID-19 patients. Thus, our objective study was to determine whether an association exists between serum concentrations of sFas and Bcl2 and COVID-19 patient mortality.Methods: This observational and prospective study of COVID-19 patients was performed in eight Intensive Care Units (ICU) from Canary Islands (Spain). Serum levels of sFas and Bcl2 at ICU admission were determined. Mortality at 30 days was the end-point study.Results: Surviving patients (n = 42) compared to non-surviving (n = 11) had lower APACHE-II (p < 0.001), lower SOFA (p = 0.004), lower serum sFas levels (p = 0.001) and higher serum Bcl2 levels (p < 0.001). Logistic regression showed an association between high serum sFas levels and mortality after controlling for APACHE-II (OR = 1.004; 95% CI = 1.101-1.007; p = 0.01) or SOFA (OR = 1.003; 95% CI = 1.101-1.106; p = 0.004), and between low serum Bcl2 levels and mortality after controlling for APACHE-II (OR = 0.927; 95% CI = 0.873-0.984; p = 0.01) or SOFA (OR = 0.949; 95% CI = 0.913-0.987; p = 0.01).Conclusions: Thus, to the best of our knowledge, this is the first study reporting blood levels of sFas and Bcl2 in COVID-19 patients and its association with mortality.
Assuntos
COVID-19/sangue , COVID-19/mortalidade , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Receptor fas/sangue , Idoso , Área Sob a Curva , Biomarcadores/sangue , Feminino , Humanos , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
OBJECTIVE: Lung cancer is one of the most prevalent cancers and the leading cause of cancer-related deaths worldwide. MicroRNAs regulate more than 60% of human genes, including tumor suppressor genes and oncogenes. Accordingly, they can affect cancer risk. This study aimed to evaluate the role of serum miR-148a as a non-invasive biomarker in non-small cell lung cancer (NSCLC) patients and to assess the correlation between miR-148a and Bcl-2, as one of its target proteins. MATERIALS AND METHODS: A total of 50 newly diagnosed NSCLC cases and 30 apparently healthy controls were recruited in this study. MiR-148a level was measured by TaqMan- Real time RT-PCR assay and Bcl-2 level was measured by ELISA. RESULTS: Significant lower expression of serum miR-148a and higher serum Bcl-2 levels were observed in NSCLC patients as compared to the control group (p.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/sangue , Neoplasias Pulmonares/sangue , MicroRNAs/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Adulto , Idoso , Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/patologia , Estudos de Casos e Controles , Egito , Feminino , Humanos , Masculino , Gradação de Tumores , Estadiamento de NeoplasiasRESUMO
INTRODUCTION AND GOAL: There is scarce and contradictory data on B-cell lymphoma 2 (Bcl2), member of the Bcl-2 antiapoptotic molecules family of intrinsic apoptosis pathway, in ischemic stroke patients. The objective of this study was to determine whether there is an association between blood Bcl2 concentrations and mortality of ischemic stroke patients. MATERIAL AND METHODS: Five Intensive Care Units participated in this prospective and observational study of patients with severe malignant middle cerebral artery infarction (MMCAI). Severe MMCAI was diagnosed when acute infarction was present in 50% or more of said region and with a Glasgow Coma Scale (GCS) score of less than 9 points. Serum samples were collected at the time of MMCAI diagnosis. FINDINGS: Higher serum Bcl2 concentrations (p = 0.001), lower platelet count (p = 0.01) and lower GCS (p = 0.002) were found in non-survivors (n = 28) than in MMCAI survivors (n = 28). Serum Bcl2 levels had an area under the curve for mortality prediction of 75% (95% CI = 62%-88%; p < 0.001). Patients with serum Bcl2 levels > 43.6 ng/mL had higher mortality rate according to Kaplan-Meier analysis (Hazard ratio=10.0; 95% CI = 3.4-29.5; p < 0.001). Multiple logistic regression showed an association between serum Bcl2 and mortality at 30 days (OR = 1.041; 95% CI = 1.006-1.077; p = 0.02) controlling for GCS and platelet count. CONCLUSIONS: This study reports for the first time the higher blood Bcl2 concentrations in non-surviving ischemic stroke patients than in survivors and the association between elevated blood Bcl2 and mortality in ischemic stroke patients.
Assuntos
Infarto da Artéria Cerebral Média/sangue , AVC Isquêmico/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Idoso , Biomarcadores/sangue , Feminino , Humanos , Infarto da Artéria Cerebral Média/diagnóstico , Infarto da Artéria Cerebral Média/mortalidade , AVC Isquêmico/diagnóstico , AVC Isquêmico/mortalidade , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Medição de Risco , Fatores de Risco , Espanha , Regulação para CimaRESUMO
miRNAs have been pointed to play critical role in the development of congenital heart disease (CHD). miRNA-375-3p (miR-375-3p) was involved in cardiac dysfunction and cardiogenesis. However, no prior study had established a therapeutic role of miR-375-3p in CHD. We intended to investigate the effect and mechanism of miR-375-3p on apoptosis in hypoxic cardiomyocytes in vitro. Expression of miR-375-3p, forkhead box P1 (FOXP1) and Bcl2 like protein 2 (Bcl2l2) was detected using real-time quantitative PCR and western blot. Apoptosis was measured with MTT assay, flow cytometry and caspase-3 activity assay. The potential target binding between miR-375-3p and FOXP1/Bcl2l2 was predicted on DianaTools, and was validated by luciferase reporter assay and RNA pull-down assay. As a result, miR-375-3p was upregulated and FOXP1/Bcl2l2 was downregulated in maternal serum of women with fetal CHD and hypoxia-induced rat cardiomyocyte h9c2 cells. Hypoxia induced apoptosis rate elevation, caspase-3 activity promotion and viability inhibition in h9c2 cells; overexpression of miR-375-3p promoted, whereas knockdown of miR-375-3p antagonized hypoxia-induced effects in h9c2 cells. In addition, miR-375-3p was validated to negatively regulate FOXP1 and Bcl2l2 expression through target binding, and silencing of FOXP1 and Bcl2l2 could independently abate the anti-apoptosis role of miR-375-3p knockdown in hypoxic h9c2 cells. Collectively, blocking miR-375-3p suppressed hypoxia-evoked apoptosis of cardiomyocytes by targeting and upregulating FOXP1 and Bcl2l2. Our results might suggest maternal serum miR-375-3p as a potential biomarker for prenatal detection of fetal CHD.
Assuntos
Fatores de Transcrição Forkhead/sangue , Cardiopatias Congênitas/sangue , MicroRNAs/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Repressoras/sangue , Animais , Apoptose/genética , Biomarcadores/sangue , Caspase 3/genética , Hipóxia Celular/genética , Modelos Animais de Doenças , Regulação da Expressão Gênica/genética , Cardiopatias Congênitas/genética , Cardiopatias Congênitas/patologia , Humanos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , RatosRESUMO
The utility of circular RNAs (circRNAs) as molecular biomarkers has recently emerged. However, only a handful of them have already been studied in colorectal cancer (CRC). The purpose of this study was to identify new circRNAs deriving from BCL2L12, a member of the BCL2 apoptosis-related family, and investigate their potential as biomarkers in CRC. Total RNA extracts from CRC cell lines and tissue samples were reversely transcribed. By combining PCR with divergent primers and nested PCR followed by Sanger sequencing, we were able to discover two BCL2L12 circRNAs. Subsequently, bioinformatical tools were used to predict the interactions of these circRNAs with microRNAs (miRNAs) and RNA-binding proteins (RBPs). Following a PCR-based pre-amplification, real-time qPCR was carried out for the quantification of each circRNA in CRC samples and cell lines. Biostatistical analysis was used to assess their potential prognostic value in CRC. Both novel BCL2L12 circRNAs likely interact with particular miRNAs and RBPs. Interestingly, circ-BCL2L12-2 expression is inversely associated with TNM stage, while circ-BCL2L12-1 overexpression is associated with shorter overall survival in CRC, particularly among TNM stage II patients. Overall, we identified two novel BCL2L12 circRNAs, one of which can further stratify TNM stage II patients into two subgroups with substantially distinct prognosis.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Colorretais/genética , Proteínas Musculares/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Circular/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/sangue , Neoplasias Colorretais/patologia , Biologia Computacional , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , MicroRNAs/sangue , Pessoa de Meia-Idade , Proteínas Musculares/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , RNA Circular/sangue , Proteínas de Ligação a RNA/sangue , Proteínas de Ligação a RNA/genéticaRESUMO
OBJECTIVE: Acute cerebral infarction (ACI) is the most common type of acute cerebrovascular disease so far, and its incidence rate has been increasing in recent years. At present, the methods of diagnosing ACI in clinic are extremely complicated, and an effective index that can effectively diagnose ACI is urgently needed in clinic. This study is designed to investigate the clinical significance of Follistatin-like protein 1 (FSTL1), Bax and Bcl-2 in ACI. PATIENTS AND METHODS: A total of 84 cases of ACI patients admitted to our hospital from September 2017 to September 2019 and 90 cases of healthy subjects undergoing physical examination at the same time were selected as the research objects for prospective analysis. The concentrations of FSTL1, Bax and Bcl-2 in the peripheral blood of objects in the two groups were detected to analyze the diagnostic value of FSTL1, Bax and Bcl-2 for ACI, and the correlation of FSTL 1, Bax and Bcl-2 with the infarct size, treatment method and hemorrhagic transformation. Another 20 SD rats were purchased, among which 10 rats were randomly selected for ACI modeling. FSTL1 concentration, Bax and Bcl-2 protein expression in brain tissues of ACI rats and normal rats were detected. RESULTS: FSTL1 and Bax in peripheral blood of ACI patients were higher than those of healthy subjects (p<0.050), and Bcl-2 was lower than those of healthy subjects (p<0.050). It was detected that FSTL1, Bax and Bcl-2 had good diagnostic value for patients with ACI (p<0.001). FSTL1 and Bax decreased while Bcl-2 increased in patients treated with thrombolytic therapy (p<0.050). And FSTL1, Bax and Bcl-2 were closely related to infarct size and hemorrhagic transformation (p<0.050). Logistic regression analysis showed that NIHSS score, atrial fibrillation, infarct volume, FSTL1 and Bax were independent risk factors affecting hemorrhagic transformation in ACI patients (p<0.050), and Bcl-2 was an independent protective factor affecting hemorrhagic transformation in ACI patients (p<0.050). The concentration of FSTL1 and the expression of Bax protein in rat brain tissue were also higher than that in normal rats, while Bcl-2 was lower than that in normal rats (p < 0.001). CONCLUSIONS: FSTL1, Bax and Bcl-2 are involved in the occurrence and development of ACI and are closely related to the hemorrhagic transformation of patients. The mechanism by which FSTL1 promotes the occurrence of ACI might be related to promoting the occurrence of inflammatory responses in the brain tissue of patients or accelerating the apoptosis of neurons.
Assuntos
Hemorragia Cerebral/tratamento farmacológico , Infarto Cerebral/tratamento farmacológico , Terapia Trombolítica , Doença Aguda , Animais , Hemorragia Cerebral/sangue , Infarto Cerebral/sangue , Proteínas Relacionadas à Folistatina/sangue , Humanos , Modelos Logísticos , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Ratos Sprague-Dawley , Proteína X Associada a bcl-2/sangueRESUMO
Dysregulation of BCL2 is a pathophysiology observed in haematological malignancies. For implementation of available treatment-options it is preferred to know the relative quantification of BCL2 mRNA with appropriate reference genes. For the choice of reference genes-(i) Reference Genes were selected by assessing variation of >60,000 genes from 4 RNA-seq datasets of haematological malignancies followed by filtering based on their GO biological process annotations and proximity of their chromosomal locations to known disease translocations. Selected genes were experimentally validated across various haematological malignancy samples followed by stability comparison using geNorm, NormFinder, BestKeeper and RefFinder. (ii) 43 commonly used Reference Genes were obtained from literature through extensive systematic review. Levels of BCL2 mRNA was assessed by qPCR normalized either by novel reference genes from this study or GAPDH, the most cited reference gene in literature and compared. The analysis showed PTCD2, PPP1R3B and FBXW9 to be the most unregulated genes across lymph-nodes, bone marrow and PBMC samples unlike the Reference Genes used in literature. BCL2 mRNA level shows a consistent higher expression in haematological malignancy patients when normalized by these novel Reference Genes as opposed to GAPDH, the most cited Reference Gene. These reference genes should also be applicable in qPCR platforms using Taqman probes and other model systems including cell lines and rodent models. Absence of sample from healthy-normal individual in diagnostic cases call for careful selection of Reference Genes for relative quantification of a biomarker by qPCR.BCL2 can be used as molecular diagnostics only if normalized with a set of reference genes with stable yet low levels of expression across different types of haematological malignancies.
Assuntos
Biomarcadores Tumorais/isolamento & purificação , Neoplasias Hematológicas/diagnóstico , Proteínas Proto-Oncogênicas c-bcl-2/isolamento & purificação , RNA Mensageiro/isolamento & purificação , RNA-Seq/normas , Animais , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Medula Óssea/patologia , Linhagem Celular Tumoral , Conjuntos de Dados como Assunto , Modelos Animais de Doenças , Estudos de Viabilidade , Regulação Neoplásica da Expressão Gênica , Genes Essenciais , Neoplasias Hematológicas/sangue , Neoplasias Hematológicas/genética , Neoplasias Hematológicas/patologia , Humanos , Leucócitos Mononucleares , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Mensageiro/sangue , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Padrões de ReferênciaRESUMO
INTRODUCTION: Histopathological examination and immunohistochemistry (IHC) with a crucial role of CD10 expression remain a standard diagnostic tool in follicular lymphoma (FL). The results of IHC CD10 detection with different primary antibodies are not fully reproducible, but some reports show that flow cytometry (FCM) can be a reliable method of CD10 identification. METHODS: The aim of the study was to compare results of CD10 expression in FL by IHC and FCM including immunophenotypic features in the context of the BCL2 and BCL6 alterations. RESULTS: Out of 76 histopathologically diagnosed FL, a group of 25 cases had simultaneously FCM. Immunohistochemically 77.6% of cases were CD10-positive with comparable and reproducible results to FCM. Differences between the FCM expression of CD5/CD10/CD11c/CD25/CD43 and BCL2 overexpression (BCL2(+)higher ) correlated with the BCL2 and BCL6 rearrangements (R) status. Lack of CD10 expression corresponded with the absence of BCL2R and higher MUM1 expression by IHC results but had no clinical impact on the long-time outcomes. CONCLUSIONS: Immunohistochemistry staining is a comparable method to FCM assessment in the evaluation of CD10 expression and diagnosis of FL. Fine-needle aspiration biopsy/FCM (FNAB/FCM) could be a useful tool for verifying FL diagnosis and CD10 detection. Despite its heterogeneity, FL has a characteristic immunophenotype. BCL2R and BCL6R FL cases differ mainly in levels of BCL2 and CD10 with CD43 co-expression; BCL2(+)higher by FCM correlates with BCL2R. Moreover, FNAB plays an important role in material provision for supportive karyotyping and BCL2R, BCL6R assessed by FISH.
Assuntos
Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica , Rearranjo Gênico , Imuno-Histoquímica , Linfoma Folicular , Neprilisina , Proteínas Proto-Oncogênicas c-bcl-2 , Proteínas Proto-Oncogênicas c-bcl-6 , Feminino , Humanos , Linfoma Folicular/sangue , Linfoma Folicular/diagnóstico , Linfoma Folicular/genética , Linfoma Folicular/patologia , Masculino , Pessoa de Meia-Idade , Neprilisina/sangue , Neprilisina/genética , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-6/sangue , Proteínas Proto-Oncogênicas c-bcl-6/genéticaRESUMO
Autism is a neurodevelopmental disorder that is recognized by stereotypic and repetitive behaviors after 2 years of old. Dysregulation of the immune system, especially inflammation which is mostly regulated by IL-6, imposes a deficit in CNS development. Along with this crucial biomarker, researchers have proposed BCL-2, micro RNA-23a-3p (miR-23a-3p), miR-181b-5p as other probable biomarkers involved in inflammation and apoptosis. The aim of the study was to evaluate the alteration in the expression of these biomarkers in a group of autism spectrum disorder (ASD) children. Peripheral blood mononuclear cells (PBMCs) were obtained from 37 autistic patients. After RNA extraction with precipitation method, the Syber green qReal-time Polymerase Chain Reaction (PCR) was performed in order to evaluate the possible alteration in the expression of IL-6, BCL-2, miR-181b-5p, and miR-23a-3p. The results were compared with healthy controls. IL-6 was significantly upregulated in ASD patients (p=0.003). On the other hand, miR-23a was upregulated and BCL-2 downregulated in ASD patients but the changes were not significant. In initial evaluations, expression changes of miR-181b-5p were not statistically significant. However, when Patients were divided into two groups of upregulated and downregulated, re-evaluation showed that both up- (p=0.005) and down-regulation (p=0.004) (i.e. changes regardless of the direction) of miR-181b were significant in autistic children. IL-6 and miR-181b-5p can have proper diagnostic values and are reliable biomarkers with high sensitivity and specificity. On the other hand, PBMC can be utilized for such studies and also evaluation of patients' condition instead of brain tissue as it is less accessible.
Assuntos
Transtorno do Espectro Autista/sangue , Biomarcadores/sangue , Interleucina-6/sangue , MicroRNAs/sangue , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Adolescente , Transtorno do Espectro Autista/diagnóstico , Transtorno do Espectro Autista/imunologia , Criança , Pré-Escolar , Feminino , Humanos , Interleucina-6/imunologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Masculino , MicroRNAs/imunologia , Proteínas Proto-Oncogênicas c-bcl-2/imunologiaRESUMO
BACKGROUND: Tumor cells with a mesenchymal phenotype and/or cancer stem-like cells (CSCs) are known to contribute to metastasis and drug resistance. Circulating tumor cells (CTCs) undergoing epithelial-mesenchymal transition (EMT) and CTCs reflecting a dedifferentiated CSC phenotype may not be detected using only an anti-EpCAM antibody to capture them. We used an antibody-independent CTC enrichment platform, ApoStream®, which does not rely on any antibody, including anti-EpCAM, to capture EMT- and CSC-CTCs in breast cancer patients who received neoadjuvant chemotherapy and correlated them to pathological complete response (pCR). METHODS: Blood samples from newly diagnosed breast cancer patients were prospectively collected before neoadjuvant chemotherapy (T0), after chemotherapy but before surgery (T1), and after surgery (T2) and processed using ApoStream. CTCs detected were stained with additional markers to define 3 CTC subsets with the following phenotypes: epithelial CTCs (CK+, EpCAM+ or E-cadherin+), EMT-CTCs (ß-catenin+ or vimentin+), and CSC-CTCs (CD44+ and CD24low). RESULTS: We enrolled 55 patients, 47 of which had data for analysis. EMT-CTCs were detected in 57%, 62%, and 72% and CSC-CTCs in 9%, 22%, and 19% at the T0, T1, and T2 time points, respectively. Counts of epithelial (P = 0.225) and EMT (P = 0.522) phenotypes of CTCs at T0 did not significantly predict pCR. Moreover, no correlation between CTC count change and pCR was demonstrated. CONCLUSIONS: ApoStream was successful in detecting EMT-CTCs among patients after neoadjuvant chemotherapy. However, EMT-/CSC-CTC counts did not correlate with pCR. Due to the small sample size and heterogeneity of this patient population, further study in a larger cohort of molecularly homogeneous patients is warranted.
Assuntos
Neoplasias da Mama/sangue , Caderinas/sangue , Molécula de Adesão da Célula Epitelial/sangue , Células Neoplásicas Circulantes/metabolismo , Adulto , Idoso , Antígenos de Neoplasias/sangue , Biomarcadores Tumorais/sangue , Neoplasias da Mama/classificação , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Contagem de Células , Linhagem Celular Tumoral , Transição Epitelial-Mesenquimal/genética , Feminino , Humanos , Pessoa de Meia-Idade , Terapia Neoadjuvante , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Vimentina/sangueRESUMO
BCL2 and BAX genes are a group of signalling inducer and inhibitor genes playing a key role in the process of cellular physiological death (apoptosis). These genes, through the JAK/STAT signalling pathway, affect different cytokines on cell function and subsequently lead to the pathophysiology of diseases, especially autoimmune diseases. In addition, altering the methylation of genes can affect their expression. Since the aetiology and pathology of Behcet's disease is not fully understood, the aim of this study was to determine the methylation pattern of BCL2 and BAX genes in patients with Behcet's disease and compare it with those of control group. This was a case-control study on 51 patients with Behcet and 61 control subjects. Blood samples were received from all subjects. Subsequently, the peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll method and the methylation of the sites was investigated using quantitative methylation specific PCR (qMS-PCR) technique after extraction of DNA by salting out method and its examination with Nano drop. The results of methylation and expression of Bax gene suggest that the methylation level in the patient group significantly increased compared to the healthy individuals (p-value < .05). Furthermore, the results related to Bax gene expression revealed that the mean of gene expression in the patient group has decreased compared to the healthy group, and this decrease was statistically significant (p-value < .05). The rate of expression and methylation of Bcl2 did not indicate any change in the two patient and healthy groups. Given the results of this study, it can be guessed that perhaps DNA methylation is involved in certain conditions of the disease and it may result in regulation of the expression of the involved genes such as Bax gene, in the pathogenesis of the disease.
Assuntos
Síndrome de Behçet/sangue , Metilação de DNA/genética , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteína X Associada a bcl-2/sangue , Adulto , Apoptose/genética , Síndrome de Behçet/genética , Síndrome de Behçet/patologia , Feminino , Regulação da Expressão Gênica/genética , Humanos , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2/genéticaRESUMO
The potential toxicity of low-dose benzene exposure to human health has received attention, but the mechanisms of low-dose benzene-induced hematotoxicity remain largely unknown. The purpose of our study was to investigate the relationships between lncRNAVNN3 expression with benzene-induced autophagy and apoptosis in control and benzene-exposed workers. Seventy benzene-exposed workers and seventy non-benzene-exposed healthy workers were recruited. The expression of lncRNAVNN3, serum autophagy-associated and apoptosis-associated proteins were evaluated, and the relationship among them were also analysed. Furthermore, the mechanism of lncRNAVNN3 on autophagy and apoptosis induced by benzene metabolite (1, 4-benzoquinone, 1, 4-BQ) was investigated in vitro. The results showed that the expression of lncRNAVNN3 increased in benzene-exposed workers (pâ¯<â¯0.05). A positive correlation was found between lncRNAVNN3, serum autophagy-associated and apoptosis-associated proteins. In addition, we found that the knockdown of lncRNAVNN3 reduced phosphorylation of beclin1 and Bcl-2, which mediated 1, 4-benzoquinone-induced autophagy and apoptosis. Overall, lncRNAVNN3 mediated 1, 4-benzoquinone-induced autophagy and apoptosis though regulating phosphorylation of beclin1 and Bcl-2, suggesting that lncRNAVNN3 might be a novel early sensitive biomarker of benzene-induced hematotoxicity.
Assuntos
Poluentes Ocupacionais do Ar/toxicidade , Amidoidrolases/metabolismo , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Benzeno/toxicidade , Moléculas de Adesão Celular/metabolismo , Exposição Ocupacional/efeitos adversos , RNA Longo não Codificante/metabolismo , Poluentes Ocupacionais do Ar/sangue , Poluentes Ocupacionais do Ar/urina , Proteína Beclina-1/sangue , Benzeno/metabolismo , Biomarcadores/sangue , Estudos de Casos e Controles , Linhagem Celular , Proteínas Ligadas por GPI/metabolismo , Humanos , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Linfócitos/patologia , Exposição Ocupacional/análise , Proteínas Proto-Oncogênicas c-bcl-2/sangueRESUMO
This study aimed to explore the effect of Sca-1+ bone marrow stromal stem cells (BMSCs) on lung ischemia reperfusion injury in mice. Five healthy Sprague-Dawley rats were selected to isolate and purify their Sca-1+ BMSCs using a Sca-1+ magnetic sorting kit in conjunction with whole bone marrow culture. In addition, 21 male C57BL/6J mice were divided into 3 groups (7 mice in each group), namely sham group (group A), I/R group (group B) and BMSCs group (group C). A pulmonary ischemia reperfusion injury model was established by ligating the left pulmonary portal vessel for 60 min and reperfusion for 240 min, after which the right pulmonary portal vessel was blocked to measure arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2). Subsequently, the mice were sacrificed to determine their superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and myeloperoxidase (MPO) activity in the lung tissue. The histological changes were observed by light microscopy, while an enzyme-linked immunosorbent assay (ELISA) was used to detect the changes in plasma expressions of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in the mice. In addition, plasma expressions of TNF-α and B-cell lymphoma-2 (bcl-2) in the mice were detected by immunohistochemistry, while the apoptosis of transplanted lung cells was detected by a TdT-mediated dUTP Nick-End Labeling (TUNEL) method. Compared with group A, group B showed a decreased level of PaO2 and SOD activity but an increased level of MDA content and MPO activity (P less than 0.01), indicating that group B had significant ischemia reperfusion injury compared to group A. In conclusion, BMSCs significantly reduced lung ischemia-reperfusion injury and improved lung function through their anti-oxidation, anti-inflammatory and anti-apoptosis properties.
Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Traumatismo por Reperfusão/terapia , Animais , Apoptose , Interleucina-10/sangue , Pulmão , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Peroxidase/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Fator de Necrose Tumoral alfa/sangueRESUMO
We investigated protein profiles specific to vascular lesions mimicking Kaposi sarcoma (KS), based on stepwise morphogenesis progression of KS. We surveyed 26 tumor-associated proteins in 130 cases, comprising 39 benign vascular lesions (BG), 14 hemangioendotheliomas (HE), 37 KS, and 40 angiosarcomas (AS), by immunohistochemistry. The dominant proteins in KS were HHV8, lymphatic markers, Rb, phosphorylated Rb, VEGF, and galectin-3. Aberrant expression of p53, inactivation of cell cycle inhibitors, loss of beta-catenin, and increased VEGFR1 were more frequent in AS. HE had the lowest Ki-67 index, and the inactivation rates of cell cycle inhibitors in HE were between those of AS and BG/KS. Protein expression patterns in BG and KS were similar. Clustering analysis showed that the 130 cases were divided into three clusters: AS-rich, BG-rich, and KS-rich clusters. The AS-rich cluster was characterized by high caveolin-1 positivity, abnormal p53, high Ki-67 index, and inactivated p27. The KS-rich cluster shared the features of KS, and the BG-rich group had high positive expression rates of galectin-3 and low bcl2 expression. In conclusion, although the rate was different, AS and HE tended to have less cell cycle marker expression than KS, and features of BG and activated KS cell signaling were similar.