RESUMO
SUMMARY: Statins inhibit cholesterol synthesis, but also have other pleiotropic effects. There are indications that they affect macrophage survival trough the regulation of apoptosis. We analyzed 50 samples of aortic wall, selected based on statins in patients' therapy (n=25, Th-S group) or statin-free therapy (n=25, Th-nonS group). Each group had 5 samples of healthy aortic tissue, 10 samples of mild and 10 samples of severe atherosclerotic changes in aortic wall. Tissue was stained with hematoxylin-eosin and immunohistochemical methods (anti-Bcl-2 antibody). Presence of Bcl2-positive macrophages (Bcl-2+ MP) was determined semiquantitatively, and data were processed in Microsoft Excell and IMB SPSS 23 Statistics. 60 % of patients in the Th-S group had a mild increase of Bcl-2+ MP The use of statins leads to a significantly more frequent increase in Bcl2+ macrophages in the intima of the healthy aortic tissue. Analysis of all aortic samples with pathohistological diagnosis showed that statin therapy was statistically significantly more often leading to a markedly increased presence of Bcl-2+ MP. In the media, all samples of the Th-S group have a mild increase of Bcl-2+ MP, and in adventitia 40 % of patients. The use of statins more often leads to a markedly increased presence of Bcl-2+ MP in aortic tissue with diagnosed mild and severe atherosclerosis. In samples of severe atherosclerosis, statins lead to a markedly increased presence of Bcl-2+ MP in the parts of the plaque towards the intima and towards the media. Statins lead to an increased presence of Bcl-2+ macrophages, prolong their life, both in healthy and atherosclerotic altered aortic tissue. This indicates potentiation of inflammation and damage to the aortic wall, and calls into question the positive effect of statins on the aortic wall with atherosclerosis.
RESUMEN: Las estatinas inhiben la síntesis de colesterol, pero también tienen otros efectos pleiotrópicos. Hay indicios de que afectan la supervivencia de los macrófagos a través de la regulación de la apoptosis.Se analizaron 50 muestras de pared aórtica, seleccionadas en base a estatinas en tratamiento de pacientes (n=25, grupo Th-S) o en tratamiento libre de estatinas (n=25, grupo Th- nonS). Cada grupo tenía 5 muestras de tejido aórtico sano, 10 muestras de cambios ateroscleróticos leves y 10 muestras de cambios ateroscleróticos severos en la pared aórtica. El tejido se tiñó con hematoxilina-eosina y métodos inmunohistoquímicos (anticuerpo anti-Bcl-2). La presencia de macrófagos positivos para Bcl2 (Bcl- 2+ MP) se determinó semicuantitativamente y los datos se procesaron en Microsoft Excell e IMB SPSS 23 Statistics. El 60 % de los pacientes del grupo Th-S tuvo un aumento leve de Bcl-2+ MP. El uso de estatinas conduce a un aumento significativamente más frecuente de macrófagos Bcl2+ en la íntima del tejido aórtico sano. El análisis de todas las muestras aórticas con diagnóstico anatomopatológico mostró que la terapia con estatinas fue significativamente más frecuente desde el punto de vista estadístico, lo que condujo a una presencia marcadamente mayor de Bcl-2+ MP. En los medios, todas las muestras del grupo Th-S tienen un leve aumento de Bcl-2+ MP, y en adventicia en el 40 % de los pacientes. El uso de estatinas con mayor frecuencia conduce a una presencia marcadamente mayor de MP Bcl-2+ en el tejido aórtico con aterosclerosis leve y grave diagnosticada. En muestras de aterosclerosis severa, las estatinas conducen a una presencia aumentada de Bcl-2+ MP en las partes de la placa hacia la íntima y hacia la media. Las estatinas conducen a una mayor presencia de macrófagos Bcl-2+, prolongan su vida, tanto en tejido aórtico sano como aterosclerótico alterado. Esto indica la potenciación de la inflamación y el daño a la pared aórtica y pone en duda el efecto positivo de las estatinas en la pared aórtica con aterosclerosis.
Assuntos
Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Aterosclerose/metabolismo , Aorta/efeitos dos fármacos , Fatores de Risco , Apoptose/efeitos dos fármacos , Medição de Risco , Genes bcl-2/fisiologia , Aterosclerose/tratamento farmacológico , Proteína bcl-X/metabolismo , Placa Aterosclerótica , Macrófagos/efeitos dos fármacosRESUMO
ABSTRACT Background: Older patients with acute myeloid leukemia are particularly difficult to treat, as they have a high risk of comorbidities, poor performance status and less tolerability to chemotherapy, as well as a more aggressive disease biology, responsible for the resistance to treatment. There is a need to explore novel therapeutic agents that are more effective and tolerable. Venetoclax, a BCL-2 inhibitor is a promising agent, as BCL-2 overexpression is present in 84% of acute myeloid leukemia patients at diagnosis and 95% of patients at relapse and has been associated with leukemia cell survival, chemotherapy resistance and poor prognosis. Objective: To review the available data about venetoclax in acute myeloid leukemia and how it can influence the treatment in older patients. Methods: Using the Pubmed database, we selected 29 articles published within the last 15 years, considering preclinical and clinical trials and review studies that combined venetoclax with acute myeloid leukemia. Results: Venetoclax has demonstrated promising results in preclinical and clinical trials, especially in patients with poor prognosis and the IDH mutation, with an excellent side-effect profile. However, resistance seems to develop rapidly with venetoclax monotherapy, because of antiapoptotic escape mechanisms. Conclusions: While the results with the use of venetoclax seem encouraging, it is not likely that targeting a single pathway will result in long-term disease control. The solution includes the use of combined therapy to block resistance mechanisms and enhance apoptosis, by reducing MCL-1, increasing BIM or inhibiting the complex IV in the mitochondria.
Assuntos
Leucemia Mieloide Aguda , Genes bcl-2 , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3 , Terapia de Alvo Molecular , Azacitidina/uso terapêutico , Decitabina/uso terapêuticoRESUMO
Abstract Objective: Myofibroblasts have been associated with the development of several pathologic fibrotic conditions. This longitudinal study aims to assess the proliferative and antiapoptotic effects of cyclosporin, nifedipine and phenytoin on gingival connective tissue cells of nonhuman primate, as well as to analyze a possible role of myofibroblasts in gingival overgrowth. Materials and Methods: Gingival samples from the right superior canine area were obtained from 12 male monkeys ( Sapajus spp ) to comprise the control group. After one week, the animals were randomly assigned to three groups, which received daily oral doses of cyclosporin, nifedipine or phenytoin for 120 days. Gingival samples were collected from the left superior canine area of two animals of each group at 52 and 120 days. Histological sections were stained with hematoxylin and eosin, and immunoreacted against α-SMA, Ki- 67 and bcl-2. Results: α-SMA immunoreaction was negative in the control and experimental groups. Similarly, no difference between groups concerning immunostaining against Ki-67 and bcl-2 was observed in connective tissue cells. Conclusion: Based on this methodology, it may be concluded that gingival overgrowths induced by cyclosporin, nifedipine and phenytoin are not associated with neither myofibroblast transdifferentiation, proliferation nor apoptosis of gingival connective cells in monkeys.
Assuntos
Animais , Masculino , Fenitoína/farmacologia , Nifedipino/farmacologia , Ciclosporina/farmacologia , Transdiferenciação Celular/efeitos dos fármacos , Miofibroblastos/efeitos dos fármacos , Gengiva/citologia , Biópsia , Imuno-Histoquímica , Distribuição Aleatória , Estudos Longitudinais , Actinas/análise , Haplorrinos , Apoptose/efeitos dos fármacos , Crescimento Excessivo da Gengiva/induzido quimicamente , Crescimento Excessivo da Gengiva/patologia , Antígeno Ki-67/análise , Antígeno Ki-67/efeitos dos fármacos , Genes bcl-2/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Miofibroblastos/citologia , Gengiva/efeitos dos fármacosRESUMO
Abstract Purpose: To investigate the role of the exogenous supply of adenosine triphosphate (ATP) in the expression of Bax and Bcl2L1 genes in intestinal ischemia and reperfusion (IR) in rats. Methods: The study was designed as a randomized controlled trial with a blinded assessment of the outcome. Eighteen adult male Wistar-EPM1 rats were housed under controlled temperature and light conditions (22-23°C, 12 h light/dark cycle). The animals were randomly divided into 3 groups: 1. Sham group (SG): no clamping of the superior mesenteric artery; 2. Ischemia and reperfusion group (IRG): 3. Ischemia and reperfusion plus ATP (IRG + ATP). ATP was injected in the femoral vein before and after ischemia. Afterwards, intestinal segments were appropriately removed and processed for Endothelial Cell Biology Rat RT2 Profiler PCR Array. Results: ATP promoted the upregulation of Bcl2L1 gene expression, whereas it did not have significant effects on Bax gene expression. In addition, the relation of Bax/Bcl2L1 gene expression in the IRG group was 1.39, whereas it was 0.43 in the IRG + ATP group. Bcl2L1 plays a crucial role in protecting against intestinal apoptosis after ischemia and reperfusion. Increased Bcl2L1 expression can inhibit apoptosis while decreased Bcl2L1 expression can trigger apoptosis. Conclusion: Adenosine triphosphate was associated with antiapoptotic effects on the rat intestine ischemia and reperfusion by upregulating of Bcl2L1 gene expression.
Assuntos
Animais , Masculino , Ratos , Trifosfato de Adenosina/farmacologia , Apoptose/efeitos dos fármacos , Genes bcl-2 , Proteína X Associada a bcl-2/genética , Isquemia/genética , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Distribuição Aleatória , Expressão Gênica , Regulação para Cima , Ratos Wistar , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Modelos Animais de Doenças , Proteína X Associada a bcl-2/efeitos dos fármacos , Proteína X Associada a bcl-2/metabolismo , Proteína bcl-X , Intestinos , Isquemia/complicaçõesRESUMO
Abstract Objective: This study aimed to investigate the protective effects of baicalin on myocardial infarction in rats and explore the related mechanisms. Methods: Fifty Sprague Dawley rats were randomly divided into the control, model, and low-, medium- and high-dose baicalin groups. The latter 3 groups were intraperitoneally injected with baicalin, with a dose of 12.5, 25 and 50 mg/kg, respectively. Then, the myocardial infarction model was established. The hemodynamic of rats was tested, the serum lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), prostacyclin (PGI2) and thromboxane A2 (TXA2) were determined, the myocardial superoxide dismutase (SOD) and malondialdehyde (MDA) levels were detected, and the myocardial B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X (Bax) protein expressions were determined. Results: Compared with the model group, in the high-dose baicalin group the ST segment height and LVEDP were significantly decreased (P<0.05), the LVSP was significantly increased (P<0.05), the serum LDH, CK-MB and TXA2 levels were significantly decreased (P<0.05), the PGI2 level was significantly increased (P<0.05), the myocardial SOD level was significantly increased (P<0.05), and the myocardial MDA level was significantly decreased (P<0.05); the myocardial Bcl-2 protein level was significantly increased, and the Bax protein level was significantly decreased (P<0.05). Conclusion: Baicalin has protective effects on myocardial infarction in rats. The possible mechanisms may be related to its resistance to oxidative stress, and up-regulation of Bcl-2 protein expression and down-regulation of Bax protein expression in myocardial tissue.
Assuntos
Animais , Flavonoides/farmacologia , Substâncias Protetoras/farmacologia , Infarto do Miocárdio/prevenção & controle , Valores de Referência , Superóxido Dismutase/análise , Tromboxano A2/sangue , Ensaio de Imunoadsorção Enzimática , Distribuição Aleatória , Reprodutibilidade dos Testes , Cromatografia Líquida de Alta Pressão , Epoprostenol/sangue , Resultado do Tratamento , Ratos Sprague-Dawley , Genes bcl-2 , Creatina Quinase Forma MB/sangue , Proteína X Associada a bcl-2/análise , Hemodinâmica/efeitos dos fármacos , L-Lactato Desidrogenase/sangue , Malondialdeído/análiseRESUMO
Abstract Purpose: To investigate the effect of melatonin on uterine tissue in the ovariectomized rat model. Methods: Fourty Wistar albino rats were divided into four groups for histologic and immunohistochemical examination. The rats were first numbered randomly and then randomly divided into 4 equal groups: control (group 1), torsion (group 2), torsion+detorsion (group 3) and torsion+detorsion+melatonin (group 4) groups. In addition, four Wistar albino rats were used for western blot analysis in each group. And also, malondialdehyde (MDA) levels were measured biochemically in all rats. Results: The histopathological examination of the uterine tissue in rats ovarectomized showed a degeneration in uterine glands, dilation of blood vessels in the internal layer with a thrombosis and bleeding, abnormal nucleuses and vacuolated cytoplasm above and below the nucleus. In torsion group, the apoptotic cells increased in luminal epithelium and gland cells. In the melatonin group showed that the Bcl2 negative effect on the uterine epithelium and did not lead to apoptotic cells. Conclusion: The increase in vascular endothelial growth factor expression resulted in the rearrangement of endothelial cell growth and the induction of angiogenesis.
Assuntos
Animais , Feminino , Útero/efeitos dos fármacos , Útero/patologia , Estro/efeitos dos fármacos , Genes bcl-2/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Melatonina/farmacologia , Antioxidantes/farmacologia , Imuno-Histoquímica , Ovariectomia , Distribuição Aleatória , Western Blotting , Actinas/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Malondialdeído/análiseRESUMO
ABSTRAC This article presents the results of a comprehensive analysis of the combined influence of genetic polymorphisms associated with various links of apoptosis regulation (BCL-2, CTLA-4 and APO-1/Fas) on the development of nodular goiter with autoimmune thyroiditis and thyroid adenoma in the studied population. The analysis was performed using the Multifactor Dimensionality Reduction (MDR) method by calculating the prediction potential. Graphic models of gene-gene interaction with the highest cross-validation consistency created by the MDR method showed complex "synergistic or independent" impact of polymorphic loci of the CTLA-4 (+49G/A), Fas (-1377G/A) and BCL-2 (63291411 A>G) genes on the onset of thyroid pathology in general, or its individual types (nodular goiter with autoimmune thyroiditis and thyroid adenoma) in the population of Northern Bukovyna.
RESUMEN Este artículo presenta los resultados de un análisis exhaustivo de la influencia combinada de polimorfismos genéticos asociados a diversos enlaces en la regulación de la apoptosis (BCL-2, CTLA-4 y APO-1/FAS) sobre el desarrollo de bocio nodular con tiroiditis autoinmune y adenoma tiroideo en la población estudiada. Para ello, se utilizó el método de reducción de dimensionalidad multifactorial (MDR) mediante el cálculo de los potenciales de predicción. Los modelos gráficos de interacción gen-gen con la mayor consistencia de validación cruzada creada por el método MDR mostraron un complejo impacto «sinérgico o independiente¼ de los loci polimórficos de los genes CTLA-4 (+49G/A), FAS (-1377G/A) y BCL-2 (63291411A>G) en el inicio de la patología tiroidea en general, o sus tipos individuales (bocio nodular con tiroiditis autoinmune y adenoma tiroideo) en la población de Bucovina septentrional.
Assuntos
Humanos , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Polimorfismo Genético/fisiologia , Tireoidite Autoimune/genética , Neoplasias da Glândula Tireoide/genética , Bócio Nodular/fisiopatologia , Bócio Nodular/genética , Apoptose/fisiologia , Receptor fas/análise , Genes bcl-2/genética , Redução Dimensional com Múltiplos Fatores/métodos , Abatacepte/análise , Bócio Nodular/etiologiaRESUMO
Several studies have revealed that certain naturally occurring medicinal plants inhibit the growth of various cancers. The present study was conducted to evaluate cytotoxicity and apoptotic induction potential of Myristica fragrans Houtt mace extract. The cytotoxic activity of the Myristica fragrans Houtt mace acetone extract was assayed by MTT assay on human oral epidermal carcinoma KB cell lines. KB cells were incubated with different concentration of mace extract ranging from 25 to 125 µg/mL for 24hrs. The apoptotic induction potential was also studied by the analysis of Bcl-2 protein and gene expression in mace extract incubated KB cell lines using western blotting technique and real-time polymerase chain reaction. The mace extract exhibited cytotoxicity and anticancer effect against KB cell lines and it also suppressed the growth of cancer cells, therefore growth inhibitory effect was noted in extract treated cell lines. The apoptotic potential of mace extract was accompanied by reduced gene expression of Bcl-2 compared to the untreated KB cells. The mace extract shows the cytotoxic activity and induced the apoptosis through the modulation of its target genes Bcl-2 in the KB cell lines, suggesting the potential of mace as a candidate for oral cancer chemoprevention. This can be further investigated in vivo for its anticancer potential.
Assuntos
Extratos Vegetais/análise , Células KB , Myristica/anatomia & histologia , Citotoxinas/análise , Plantas Medicinais/classificação , Preparações Farmacêuticas , Apoptose , Genes bcl-2/fisiologiaRESUMO
BACKGROUND: Glioma is the most prevalent malignant tumor in human central nervous systems. Recently, the development of resistance to radiotherapy in glioma patients markedly vitiates the therapy outcome. MiR-153-3p has been reported to be closely correlated with tumor progression, but its effect and molecular mechanism underlying radioresistance remains unclear in glioma. METHODS: The expression of miR-153-3p was determined in radioresistant glioma clinical specimens as well as glioma cell lines exposed to irradiation (IR) using quantitative real-time PCR. Cell viability, proliferation and apoptosis were then evaluated by MTT assay, colony formation assay, Flow cytometry analysis and caspase-3 activity assay in glioma cells (U87 and U251). Tumor forming was evaluated by nude mice model in vivo. TUNEL staining was used to detect cell apoptosis in nude mice model. The target genes of miR-153-3p were predicted and validated using integrated bioinformatics analysis and a luciferase reporter assay. RESULTS: Here, we found that miR-153-3p was down-regulated in radioresistant glioma clinical specimens as well as glioma cell lines (U87 and U251) exposed to IR. Enhanced expression of miR-153-3p promoted the radiosensitivity, promoted apoptosis and elevated caspase-3 activity in glioma cells in vitro, as well as the radiosensitivity in U251 cell mouse xenografs in vivo. Mechanically, B cell lymphoma-2 gene (BCL2) was identified as the direct and functional target of miR-153-3p. Moreover, restoration of BCL2 expression reversed miR-153-3p-induced increase of radiosensitivity, apoptosis and caspase-3 activity in U251 cells in vitro. In addition, clinical data indicated that the expression of miR-153-3p was significantly negatively associated with BCL2 in radioresistance of glioma samples. CONCLUSIONS: Our findings suggest that miR-153-3p is a potential target to enhance the effect of radiosensitivity on glioma cells, thus representing a new potential therapeutic target for glioma.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Tolerância a Radiação/genética , Genes bcl-2/fisiologia , MicroRNAs/efeitos da radiação , MicroRNAs/fisiologia , Glioma/genética , Fatores de Tempo , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Sobrevivência Celular/efeitos da radiação , Western Blotting , Análise de Variância , Marcação de Genes/métodos , Genes bcl-2/efeitos da radiação , Marcação In Situ das Extremidades Cortadas , MicroRNAs/análise , Linhagem Celular Tumoral , Proliferação de Células/efeitos da radiação , Caspase 3/análise , Reação em Cadeia da Polimerase em Tempo Real , Citometria de Fluxo , Glioma/radioterapiaRESUMO
The antidiabetic drug metformin has been found to have beneficial effects in various neurological disorders; however, the molecular mechanisms underlying these effects remain unclear. Here we report that metformin protects neuronal cells from quinolinic acid (QUIN)-induced excitotoxicity. For this, we pretreated N18D3 neuronal cells with metformin prior to QUIN for 24 h. We found that pretreating the cells with metformin significantly improved cell survival rate in a concentration-dependent manner and reduced apoptotic cell death, as revealed by a MTT assay and DAPI staining, respectively. Calcium imaging using fluo-4 showed that metformin (100 µM) inhibited the intracellular calcium increase that was induced by QUIN. In addition, mRNA expression of pro-apoptotic genes, p21 and Bax, was decreased and of anti-apoptotic genes, Bcl-2 and Bcl-xl, was increased with metformin treatment compared to QUIN-induced cells. The immunoreactivity of phosphorylated ERK1/2 was elevated in cells treated with metformin, indicating the ERK1/2 signaling pathway in the neuroprotective effects of metformin in QUIN-induced cell death. Collectively, our data demonstrates that metformin exerts its neuroprotective effects by inhibiting intracellular calcium increases, allowing it to regulate ERK1/2 signaling and modulate cell survival and death genes.
Assuntos
Apoptose , Cálcio , Morte Celular , Sobrevivência Celular , Genes bcl-2 , Metformina , Doenças do Sistema Nervoso , Neurônios , Neuroproteção , Fármacos Neuroprotetores , Ácido Quinolínico , RNA MensageiroRESUMO
Objective: To compare the rate of cell proliferation and expression of antiapoptotic protein Bcl-2 between drug-induced gingival overgrowth (DIGO) and clinical healthy gingiva (CHG) and to establish associations with histopathological features. Material and Methods: Twenty specimens of DIGO and 20 CHG specimens were submitted to morphological and immunohistochemical analysis by light microscopy. Cell proliferation (Ki-67) and the expression of Bcl-2 were evaluated in epithelial cells and spindle-shaped mononuclear cells of the connective tissue by establishing the labeling index (LI). Results: In epithelial tissue, the mean LI for Ki-67 was 17.2% in DIGO and 21.71% in CHG (p = 0.137). The mean LIs for Bcl-2 in epithelial tissue were 14.67% and 10.24% in DIGO and CHG, respectively (p = 0.026). In connective tissue, DIGO and CHG specimens exhibited low LIs for Ki-67 and Bcl-2, with mean values of less than 0.5% in both groups. No significant differences in the LIs for Ki-67 or Bcl-2 in epithelial tissue were observed according to the degree of collagenization, degree of vascularization and intensity of inflammatory infiltration (p > 0.05). No significant correlations were observed between the LIs for Ki-67 and Bcl-2 (p > 0.05). Conclusion: The present results suggest that the pathogenesis of DIGO does not involve increased proliferation or decreased apoptosis of fibroblasts. On the other hand, the morphological pattern of elongated epithelial cristae observed in DIGO could mainly be due to the inhibition of keratinocyte apoptosis and not to increased proliferation of these cells.
Assuntos
Proliferação de Células , Fibromatose Gengival/patologia , Genes bcl-2 , Imuno-Histoquímica/métodos , Antígeno Ki-67 , Brasil , Estatísticas não ParamétricasRESUMO
The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.
Assuntos
Humanos , Apoptose , Catalase , Sobrevivência Celular , Expressão Gênica , Genes bcl-2 , Glutationa Peroxidase , Glutationa Redutase , Mãos , Malondialdeído , Estresse Oxidativo , Próstata , Neoplasias da Próstata , Reação em Cadeia da Polimerase em Tempo Real , Venenos de Escorpião , Escorpiões , Venenos de Serpentes , Serpentes , Superóxido Dismutase , Peçonhas , Venenos de Víboras , ViperidaeRESUMO
Background: Apoptosis-related gene expression such as BCL2, and p53 has been suggested in predicting the patient response to chemo- or radiotherapy, as well as patient's survival. Methods: The aim of this study was to determine changes in BCL2 and p53 apoptosis related gene expressions in chronic lymphocytic leukemia (CLL) patients in response to different chemotherapy regimens and number of treatment courses. The study was conducted on 55 CLL patients (44 CLL and 11 CLL/SLL; small lymphocytic lymphoma) and 40 healthy individuals as control, over three-months period. The RNA was extracted by exploitation total RNA extraction kit, treated with DNAse, then cDNA was synthesized and qRT-PCR used to analyze antiapoptotic BCL2 and tumor suppresser p53 gene expressions. Results: CLL/SLL showed higher BCL2 and p53 gene expression than CLL. The patients with CLL showed three-fold increase in BCL2 gene expression compared to healthy controls (p < 0.05), and 50% decrease in p53 gene expressions (p < 0.05). BCL2 gene expression was higher, particularly, for those who were treated with higher range of treatment courses and combination of fludarabine, cyclophosphamide and rituximab (FCR) regimen. P53 gene expression reciprocally related with BCL2 and vice versa. Conclusions: In contrary to BCL2, p53 gene was extremely expressed in patients treated with chemotherapy agents, particularly after 2430 months disease duration; suggesting a late expression of p53 during advanced stages of the disease. A proportional change in BCL2 and p53 gene expression was reported with different treatment regimens; Chlorambucil (Clb) decreased and FCR regimen increased BCL2 gene expression. Higher p53 gene expression reported with the Chlorambucil + (Chlorambucil + Prednisolone) regimen (AU)
Assuntos
Humanos , Masculino , Feminino , Leucemia Linfocítica Crônica de Células B , Leucemia , Expressão Gênica , Genes p53 , Apoptose , Genes bcl-2 , QuimiorradioterapiaRESUMO
OBJECTIVE@#To investigate the process of apoptosis in lungs and liver induced by crushing hindlimbs of rat, and study the mechanism of crush injury.@*METHODS@#The rat experimental model of hindlimbs crush injury was established. The cell apoptosis in lungs and liver was detected by TUNEL assay, and the expression of Bax, Bcl-2 and caspase-3 apoptin was examined by immunohistochemistry.@*RESULTS@#Compared with the control group, the partial muscle injury of rat's hindlimbs was more serious with more apoptosis observed in lungs and liver (P < 0.05). The expression of Bax was up-regulated and Bcl-2 was down-regulated, whereas caspase-3 expression was activated (P < 0.05).@*CONCLUSION@#The cell apoptosis has increased significantly in lungs and liver after crush injury of hindlimbs in rat. The correlation factor released during tissue injury may mediate apoptosis process.
Assuntos
Animais , Ratos , Apoptose/fisiologia , Caspase 3/metabolismo , Genes bcl-2 , Membro Posterior/lesões , Imuno-Histoquímica , Fígado/fisiopatologia , Pulmão/fisiopatologia , Regulação para Cima , Proteína X Associada a bcl-2RESUMO
Lichen planus is a common immune-mediated disease that is associated with an increased risk of malignant transformation in the oral cavity. Synchronous up-regulation of COX-2 and down-regulation of Bcl-2 have been demonstrated in some malignancies. The aim of this study was to assess the correlation between COX-2 and Bcl-2 expression and their role in dysplastic changes of oral lichen planus [OLP]. This study was performed on 47 paraffin blocks with the diagnosis of OLP and 16 blocks with the diagnosis of focal fibrous hyperplasia [control group]. Immunohistochemical staining was performed using antibodies against COX-2 and Bcl-2. Spearman's rank correlation coefficient and Mann-Whitney test were used for data analysis. A significant correlation was observed between the intensity of sub-epithelial inflammation and the severity of basal cell layer degeneration [P=0.048]. Significant up-regulation of Bcl-2 and COX-2 was detected in sub-epithelial inflammatory infiltration [P<0.001, P=0,003]. The amount and intensity of Bcl-2 and COX-2 expression were significantly correlated in sub-epithelial lymphocytic infiltration [P=0.013, P=0.019]. Our findings indicated the effective role of Bcl-2 expression in decreasing the apoptosis in the inflammator y infiltrate unlike the epithelium. The significant correlation of the intensity of Bcl-2 expression in the epithelium and the sub-epithelial inflammatory infiltrate with COX-2 expression and also the correlation of the intensity of inflammation with the severity of basal layer hydropic degeneration may imply that these two markers can induce malignant transformation in the affected epithelium in an indirect manner by the continuation of inflammation and activation of carcinogenic mechanisms
Assuntos
Genes bcl-2 , Ciclo-Oxigenase 2 , Expressão GênicaRESUMO
To evaluate the mRNA expression ratio of Bcl-2/Bax both in normal and tumoral bladder tissues of patients with transitional cell carcinoma [TCC] of bladder and investigate potential correlation between this expression ratio and clinical outcome. In this experimental study, we used real time-PCR to investigate the expression of Bcl-2 and Bax both in normal and tumoral bladder tissues. The Bcl-2/Bax expression ratio was determined in tumoral bladder tissues of patients with transitional cell carcinoma of the bladder [n=40] and correlation between expression ratios and the emergence of early relapses in a follow-up of 14-30 months was examined. Relapse-free time in 14/31 patients [45.16%] with Bcl-2/Bax>1 was shorter than 9 months [range of 2-9 months] with 5.7 months average median while 17/31 patients [54.84%] with Bcl-2/Bax<1 are currently relapse-free [14-30 months]. Bcl-2 and Bax expression levels were not solely correlated with clinical outcome and progression of carcinogenesis. The mRNA expression ratio of Bcl-2/Bax in tumoral bladder tissues may serve as a significant prognostic indicator in predicting the clinical outcome in low grade non-invasive bladder cancer
Assuntos
Humanos , Masculino , Carcinoma de Células de Transição , Genes bcl-2 , Proteína X Associada a bcl-2 , Reação em Cadeia da Polimerase em Tempo Real , RNA MensageiroRESUMO
Telomerase activity increases in cancer cells. Bcl-2 is an antiapoptotic factor that its concentration grows in many cancer cells including hepatocellular carcinoma cells. In this study, an attempt was made to investigate the effects of a new synthetic compound, platinum azidothymidine [Pt-AZT] on treatment of rats with Hepatocellular Carcinoma [HCC] and to compare its effects with azidothymidine [AZT] in alteration of telomerase activity and Bcl-2 concentration in HCC. Healthy adult male Wistar rats [n=100] were randomly divided into 4 groups [A, B, C, and D]. Group A contained 25 healthy rats and was considered as the control group. Liver preneoplastic lesions were induced in remaining animals [n=75] using Solt-Farber resistant hepatocyte protocol. These animals were randomly allocated in groups B, C and D. Group B was negative control [untreated], groups C and D were treated by intraperitoneal injection [IP] of Pt-AZT [0.9 mg/kg/day] and AZT [0.3 mg/kg/day], respectively for 14 days. After the treatment period, telomerase activity and Bcl-2 concentration were determined in the rats' liver. No HCC was developed in group A, but tumors were present in all other groups. Telomerase activity and Bcl-2 concentration were significantly lower in group C compared to groups B [0.159 +/- 0.06 vs. 0.577 +/- 0.116 IU/L, p<0.001, respectively and 0.931 +/- 0.388 vs. 3.94 +/- 0.74 ng/ml, p<0.001, respectively]. Similar results were observed in comparison with group D [0.331 +/- 0.06 vs. 0.577 +/- 0.116 IU/L, p<0.001, respectively and 0.931 +/- 0.388 vs. 2.94 +/- 0.594 ng/ml, respectively]. There was a significant negative correlation between telomerase activity and Bcl-2 concentration only in untreated cancer group [p=0.034]. In this study, higher anticancer activity of Pt-AZT in comparison to AZT was demonstrated. It effectively inhibits the growth of liver tumor in rats through extending apoptosis
Assuntos
Animais de Laboratório , Platina , Zidovudina , Telomerase , Genes bcl-2 , Neoplasias Hepáticas , Neoplasias Hepáticas Experimentais , Ratos WistarRESUMO
BACKGROUND: Vascular endothelial growth factor (VEGF) is involved in the growth of new blood vessels that feed tumors and kinesin spindle protein (KSP) plays a critical role in mitosis involving in cell proliferation. Simultaneous silencing of VEGF and KSP, an attractive and viable approach in cancer, leads on restricting cancer progression. The purpose of this study is to examine the therapeutic potential of dual gene targeted siRNA cocktail on human hepatocellular carcinoma Hep3B cells. RESULTS: The predesigned siRNAs could inhibit VEGF and KSP at mRNA level. siRNA cocktail showed a further downregulation on KSP mRNA and protein levels compared to KSP-siRNA or VEGF-siRNA, but not on VEGF expression. It also exhibited greater suppression on cell proliferation as well as cell migration or invasion capabilities and induction of apoptosis in Hep3B cells than single siRNA simultaneously. This could be explained by the significant downregulation of Cyclin D1, Bcl-2 and Survivin. However, no sigificant difference in the mRNA and protein levels of ANG2, involving inhibition of angiogenesis was found in HUVECs cultured with supernatant of Hep3B cells treated with siRNA cocktail, compared to that of VEGF-siRNA. CONCLUSION: Silencing of VEGF and KSP plays a key role in inhibiting cell proliferation, migration, invasion and inducing apoptosis of Hep3B cells. Simultaneous silencing of VEGF and KSP using siRNA cocktail yields promising results for eradicating hepatocellular carcinoma cells, a new direction for liver cancer treatment.
Assuntos
Humanos , Cinesinas/genética , Apoptose/genética , Inativação Gênica , RNA Interferente Pequeno/genética , Fator A de Crescimento do Endotélio Vascular/genética , Proliferação de Células/genética , Sais de Tetrazólio , Transfecção , Inibidores de Cisteína Proteinase/metabolismo , Regulação para Baixo , Movimento Celular , Western Blotting , Cinesinas/metabolismo , Anexina A5 , Genes bcl-2 , Ciclina D1/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Linhagem Celular Tumoral , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas Inibidoras de Apoptose/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Citometria de Fluxo , Survivina , Mitose/genéticaRESUMO
<p><b>OBJECTIVE</b>To explore the expression of Bcl-2 mRNA and its effect on prognosis of patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).</p><p><b>METHODS</b>Real time quantitative PCR was used to determine the expression of Bcl-2 mRNA in 40 PGI-DLBCL patients and 17 healthy controls. The association of Bcl-2 expression with clinicopathological features and prognosis of the patients was analyzed.</p><p><b>RESULTS</b>The expression level of Bcl-2 mRNA in PGI-DLBCL patients was 1.03 ± 0.93, significantly higher than that of the controls (0.41 ± 0.21) (P < 0.05). The expression of Bcl-2 mRNA in stage IIE-IV patients (1.28 ± 1.01) was significantly higher than that in the stage I-II2 patients (0.62 ± 0.61) (P < 0.05). The expression of Bcl-2 mRNA in patients with international prognostic index (IPI) score >2 (1.95 ± 1.27) was significantly higher than those with IPI score ≤ 2 (0.86 ± 0.75)(P < 0.05). The expression of Bcl-2 mRNA in patients with complete remission (CR) (0.71 ± 0.58) was significantly lower vs. 2.42 ± 0.91 in patients with no CR (P < 0.05). Univariate analysis indicated that β2-MG, IPI score>2, the Lugano staging, and Bcl-2 mRNA expression were associated with overall survival (OS) and progression-free survival (PFS) (P < 0.05). Multivariate analysis indicated that IPI score>2 was independently associated with OS (P < 0.05), and both IPI score >2 and Bcl-2 mRNA expression were independently associated with PFS (P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of Bcl-2 mRNA in the tumor tissue of PGI-DLBCL patients is significantly higher than that in controls. PGI-DLBCL patients with higher expression of Bcl-2 have a poor chemotherapy response and inferior prognosis. IPI score >2 and higher expression of Bcl-2 mRNA are independent poor prognostic factors for PFS in PGI-DLBCL patients.</p>
Assuntos
Humanos , Intervalo Livre de Doença , Genes bcl-2 , Linfoma de Células B , Diagnóstico , Genética , Metabolismo , Linfoma Difuso de Grandes Células B , Diagnóstico , Genética , Metabolismo , Prognóstico , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , MetabolismoRESUMO
Ischemia-reperfusion injury is the tissue damage caused when blood supply returns to the tissue after a period of ischemia or lack of oxygen. Ischemia-reperfusion brain injury initiates an inflammatory response involving the expression of adhesion molecules and cytokines. Twenty-four male Wistar rats [250-300 g body wt] were used in this study. The animals were divided into four groups of 6 rats each: I: Control group that was subjected to ischemia-reperfusion, II: Ischemia-reperfusion group that was subjected to all surgical procedures, III: Drug group that received pentoxifylline [200, 400 and 600 mg/kg] 60 min before and after ischemia and IV: Vehicle group that received saline. Seventy two h after ischemia-reperfusion, the hippocampus was taken for studying the changes in bcl-2 gene expression. We used quantitative real-time PCR for the detection of bcl-2 gene expression in ischemia and drug groups and then compared them to normal samples. The results showed the gene dosage ratio of 0.66 and 1.5 for ischemia group and the drug groups, respectively. The results also showed the bcl-2 gene expression declined in ischemia group as compared to the drug group. Furthermore, we observed a significant difference in the bcl-2 gene expression between ischemia and drug groups. These findings are consistent with anti-apoptotic properties of bcl-2 gene. Furthermore this method provides a powerful tool for the investigators to study brain ischemia and respond to the treatment drugs with anti-apoptotic agents