Associação do polimorfismo genético no gene BCL2 no Acidente Vascular Cerebral Hemorrágico/Aneurisma / Association of genetic polymorphism in the BCL2 gene in/with Hemorrhagic Stroke/Aneurysm / Asociación de polimorfismo genético en el gen BCL2 en/con el accidente cerebrovascular hemorrágico/aneurisma

Objetivo: Verificar a distribuição do polimorfismo do gene BCL2 (rs1801018), em sua região codante, em pacientes portadores de Acidente Vascular Cerebral Hemorrágico (AVCh)/Aneurisma. Além de associar o presente polimorfismo as manifestações clinicas da doença. Método: O estudo foi conduzido com 158 participantes de pesquisa. Os grupos foram pareados quanto ao sexo e idade. A genotipagem foi conduzida pela técnica PCR-RFLP. Após o cálculo das frequências alélicas e genotípicas de cada grupo, foram utilizados testes estatísticos apropriados para cada tipo de comparação. O nível de significância adotado foi de 5%. Resultados: Os dados indicaram que a frequência dos genótipos apresentou uma diferença estatisticamente significante entre o grupo caso e controle, encontrando-se o genótipo ala43ala na maioria dos participantes de ambos os grupos. Conclusão: A presença do alelo mutante (trh) foi vista como um fator protetor para AVCh/aneurisma. Porém, estudos em outras populações devem ser realizados para se obter uma melhor compreensão sobre a doença AVCh/aneurisma.

Objective: Verify the distribution of the BCL2 gene polymorphism (rs1801018), located in its coding region, in patients with Hemorrhagic Stroke (HS)/Aneurysm (A). Furthermore, to associate this polymorphism with the HS/A clinical manifestations. Method: The study was conducted with 158 research participants and the groups matched by sex and age. Genotyping was done by the PCR-RFLP technique. After calculating the allele and genotype frequencies of each group, appropriate statistical tests were performed for each comparison type with the adopted significance level of 5%. Results: The data indicated that the frequency of the genotypes showed a statistically significant difference between the case and control group, and the ala43ala genotype was found in most participants in both groups. Conclusion: The presence of the mutant allele (trh) was observed as a protective factor for HS/A. However, studies in other populations should be performed to obtain a better understanding of this disease.

Objetivo: Objetivo: Verificar la distribución del polimorfismo del gen BCL2 (rs1801018), en ubicado su región de codificación, en pacientes con accidente cerebrovascular hemorrágico (ACVh)/aneurisma. Asimismo, asociar el presente polimorfismo con las manifestaciones clínicas de la enfermedad. Método: El estudio se realizó con 158 participantes, y los grupos fueron agrupados por sexo y edad. La determinación del genotipo se realizó mediante la técnica PCR-RFLP. Después de calcular las frecuencias de alelos y genotipos de cada grupo, se realizaron pruebas estadísticas apropiadas para cada tipo de comparación con el nivel de significación adoptado de 5%. Resultados: Los datos indicaron que la frecuencia de los genotipos mostró una diferencia estadísticamente significativa entre el grupo caso y el control, con el genotipo ala43ala encontrado en la mayoría de los participantes de ambos grupos. Conclusión: La presencia del alelo mutante (trh) fue visto como un factor protector para el ACVh/aneurisma. Sin embargo, se deben realizar más estudios en otras poblaciones para obtener una mejor comprensión de la enfermedad de ACVh/aneurisma.

The effects of coadministration of tilorone dihydrochloride and culture supernatants from Lactobacillus reuteri on the mouse hepatoma cell line

Purpose: This trial studies the feasibility and potential utility of stereotactic body radiation therapy in patients with unresectable liver metastasis. Aims: (1) The aim of this study is to assess the local response of the liver lesions poststereotactic body radiation therapy regarding number and size of lesions and (2) to evaluate the toxicity to organ (s) at risk. Materials and Methods: A total of 15 patients were enrolled in this study from November 2014 to October 2015. The inclusion criteria for this study were patients having 1–3 liver metastasis from any solid tumor except germ cell tumor or lymphoma with no evidence of progressive disease (PD) outside the liver. A planning four dimensional-computed tomography (CT) scan was taken. Planning target volume was generated by giving margin of 5 mm. Dose prescribed was 36 Gy in 3#. Response was defined by CT abdomen done at 3 and 6 months poststereotactic body radiation therapy as per RECIST guideline (v1.1). Results: At 3 months poststereotactic body radiation therapy, five patients had partial response, five patients had stable disease, and five patients had PD as per RECIST criteria. Out of 20 assessable lesions, 16 were controlled at 3 months poststereotactic body radiation therapy. The actuarial local control rate was 86% at 3 months and 77% at 6 months poststereotactic body radiation therapy. The median progression free survival was 7 months. Two patients experienced Grade 2 gastric toxicity and one patient experienced Grade 2 small bowel toxicity. No cases of radiation-induced liver disease were observed. Conclusions: This trial examines the feasibility of stereotactic body radiotherapy to liver metastasis in the Indian scenario. It shows excellent tolerability and is a safe therapeutic option for inoperable patients, showing good local control

Salvia miltiorrhiza bge. f. alba up-regulates Bcl-2 and inhibits apoptosis of human umbilical vein endothelial cells / 第二军医大学学报

Objective To study the influence of Salvia miltiorrhiza bge. f. alba(SMA) on apoptosis and Bcl-2 expression in human umbilical vein endothelial cell (HUVEC) in vitro. Methods HUVECs were isolated using perfusion and enzyme digestion methods, and the obtained cells were identified by morphological observation and VIII = Ag immunoreactivity examination. The cells in the exponential phase of growth were treated with H2O2 and different concentrations of SM A (high dose, 0.10 g/ml, low dose, 0.01 g/ml). The cell apoptosis was determined by flow cytometric analysis and Bcl-2 expression was examined by immunofluorescence mehtod. Results SMA significantly decreased the H2O2-induced apoptosis of HUVECs (P<0.01 in high dose group and P<0.05 in low dose group), and significantly increased the expression of Bcl-2 in high dose group(P<0.01). Conclusion SMA can inhibit H2O2-induced apoptosis of HUVEC, which might be associated with the increase of Bcl-2 expression. Salvia miltiorrhiza bge. f. alba; umbilical veins; endothelial cell; apoptosis; bcl-2 genes.

Effect of Human Bcl-2 Gene Expression on the Peripheral Atherosclerotic Lesions of Apolipoprotein E-Deficient Mouse

BACKGROUND AND OBJECTIVES: Bcl-2 protein is related to the inhibition of apoptosis via the mitochondrial pathway and Bcl-2's anti-oxidant effect. During the development of atherosclerosis, apoptosis is known to play an important role in the pathophysiologic behavior of atherosclerotic vascular disease in the medium-sized arteries. Apoptosis may be a compensatory reaction to regulate the cellular density of various tissues during the cellular proliferation process such as happens with tissue injury and during the development of atherosclerosis. The consequences of apoptosis in atherosclerosis may be related to the formation of an acellular lipid core, plaque instability and the loss of vascular wall integrity and remodeling. We sought to determine the effect of Bcl-2 gene expression on the development of primary atherosclerosis in apolipoprotein E deficient mouse, which is one of the typical animal models that are used for the development of peripheral atherosclerosis. MATERIALS AND METHODS: Bcl-2 transgenic mice were cross hybridized with apolipoprotein E deficient mice. Systemic analysis of the distribution and severity of their atherosclerotic lesions was done by dissecting microscopy, and the histological characteristics of the lesions were evaluated in normal chow-fed, 9-month-old apolipoprotein-E deficient/Bcl-2 transgenic mice (n=6) and apolipoprotein-E deficient mice (n=6). RESULTS: The distribution and severity of atherosclerotic lesions at the peripheral arteries were less in the apolipoprotein-E deficient/Bcl-2 transgenic mice. Acellular lipid core formation, destruction of the smooth muscle cell layers in the media and infiltration of inflammatory cells in the adventitia were much less in the apolipoprotein-E deficient/Bcl-2 transgenic mice. The lipid profile was similar in both groups. CONCLUSION: The effect of Bcl-2 gene expression on the peripheral atherosclerosis was related with the inhibition or the delay of atherosclerotic lesion progression, such as the reduction of amount of the acellular lipid core, maintenance of vascular smooth muscle cell integrity and the reduction of adventitial inflammation, and this was achieved regardless of serum cholesterol level.

Induction of Apoptosis by a Novel Bcl-2 Protein, Bak-like without a BH3 Domain

OBJECTIVE: In a variety of physiologic settings, cells are eliminated by apoptosis, a genetically encoded process of cellular suicide. Bak, a member of the Bcl-2 protein family, accelerates apoptosis by an unknown mechanism. Here, we describe the identification and characterization of a complementary DNA that encodes a previously unknown Bcl-2 homologue designated Bak-like. METHODS: We identified a splicing variant of Bak with a lacked BH3 domain from human full-length cDNA bank. The expression of Bak-like was examined by northern blot analysis and polymerase chain reaction. To investigate whether Bak-like might arise from alternative splicing of mRNA of Bak, Southern blot analysis was executed. Apoptosis in transfected HeLa cells was analyzed by direct counting of viable cells. We examined the location of Bak-like in individual living cells by using EGFP fusion constructs and confocal microscope. RESULTS: Bak-like cDNA coded a protein consisting of 101 amono acid, and conserved BH1 and BH2 domains like Bak but not BH3 domain. Bak-like mRNA was about 2.4kb similar to bak. Bak-like was assumed to be an alternative splicing variant of Bak and to concern with promotion of apoptosis. GFP-bak-like markedly changed its intracellular distribution, relocating within cells during apoptosis from a diffuse to a punctate pattern. CONCLUSION: Our results define a novel splicing form of the bak gene and demonstrate that this variant without a conserved BH3 domain appears to contain the BH1 and BH2 domains and the transmembrane sequence for apoptosis induction by channel-forming Bcl-2 proteins. Like Bak, Bak-like gene product primarily enhanced apoptotic cell death following an appropriate stimulus.

Expression of Bcl-2 and Bax in cis-Diamminedichloroplatinum (II)-Resistant Bladder Cancer Cell Lines / 대한비뇨기과학회지

PURPOSE: Development of drug resistance has been the major obstacle in cis-Diamminedichloroplatinum (II) (cisplatin)-based combination chemotherapy in the treatment of advanced bladder cancer for which a variety of mechanisms has been suggested. We investigated to determine the changes of expression of apoptotic regulator proteins Bcl-2 and Bax in cisplatin-resistant bladder cancer cell lines and the reversibility of chemoresistance with antisense oligonucleotide against Bcl-2. MATERIALS AND METHODS: In T24, J82, 253J, 253J-BV and HT-1376 bladder cancer cell lines, we established cisplatin-resistance using stepwise exposure to cisplatin. The changes of Bcl-2 and Bax proteins in the resistant cell lines were determined by Western blot. Then, after administration of antisense oligonucleotide targeting the Bcl-2 coding sequence to the T24, T24-R1, and T24-R2 cell lines with lipofectamine, changes of Bcl-2 expression were determined along with cisplatin cytotoxicity before and after transfection. RESULTS: We confirmed the acquisition of cisplatin resistance in all 5 cell lines as the percent increase of IC50 in each cell lines were 210%, 175%, 181%, 280% and 153%, respectively. The expression of Bcl-2 protein increased in all 5 cisplatin-resistant cell lines, while the expressions of Bax decreased in 4 of 5 cisplatin-resistant cell lines. Treatment with antisense oligonucleotide significantly enhanced the cytotoxicity of cisplatin in T24, T24-R1 and T24-R2 cell lines. CONCLUSIONS: These results suggest that the up-regulation of Bcl-2 expression as well as down-regulation of Bax expression may be one of the mechanisms of cisplatin resistance in bladder cancer cells, and antisense Bcl-2 oligonucleotide may be helpful in chemotherapy of bladder cancer by reversing cisplatin resistance.

Mechanism of Azaline B-induced Apoptosis of Ventral Prostate in Rat / 대한비뇨기과학회지

PURPOSE: Androgen deprivation triggers a sequence of events that activates apoptotic cell death of the androgen-dependent epithelial cells within the rat ventral prostate, ultimately resulting in the involution of the gland. To investigate the mechanism of azaline B-dependent apoptosis in the rat ventral prostate, the regulation of apoptosis-related genes were examined. MATERIALS AND METHODS: Azaline B was subcutaneously injected in Sprague-Dawley rat. Fas receptor(Fas), Fas ligand(FasL), bcl-2 mRNA, and protein levels were detected by RT-PCR and Western blot. Azaline B-dependent apoptosis was determined by TUNEL and DNA fragmentation assay. Transacting factor of FasL promoter was identified by DNA footprinting and DNA mobility shift assay. RESULTS: The prostate regressed after azaline B treatment in rat, and the involuted ventral prostate regenerated after testosterone pretreatment. Apoptosis of the ventral prostate was detected by TUNEL assay and apoptotic DNA fragmentation assay after azaline B treatment. The levels of Fas and FasL mRNA and protein increased after azaline B treatment. In DNase I footprinting assay with FasL promoter using nuclear extract prepared from control prostate, at least two sites were protected: SP-1 binding site at -283bp and prostate-unidentified factor(P-UF) binding site at -247bp. SP-1 binding activity vanished in the nuclear extract prepared from azaline B-treated rats. In the DNA mobility shift assay, SP-1 binding activity decreased after azaline B treatment. Bcl-2 mRNA and protein were downregulated after azaline B treatment. CONCLUSIONS: These results suggest that Fas/FasL system and Bcl-2 are important to azaline B-dependent apoptosis in rat ventral prostate and that SP-1 is related to azaline B-dependent regulation of the FasL gene.