ABSTRACT
Plasma cell disorders are a group of heterogeneous diseases originating from plasma cells, including multiple myeloma, plasma cell leukemia and light-chain amyloidosis, etc. Monoclonal plasma cells are detected in bone marrow and affected tissues, monoclonal immunoglobulin or components are detected in serum or urine, and some end-organs are injured. Plasma cell disorders accompanied by t(11;14) have unique biological characteristics and unsatisfactory response to proteasome inhibitors. With t(11;14) translocation, the expressions of cyclin D1 and anti-apoptotic protein bcl-2 are relatively high, which lead to the occurrence of plasma cell disorders and have implications for the prognosis of disease. Venetoclax is a bcl-2 inhibitor, and its single agent or combined with other drugs has achieved good efficacy in treatment of plasma cell disorders with t(11;14). This article reviews the progress of bcl-2 inhibitors in treatment of plasma cell disorders.
ABSTRACT
Objective:To investigate the expressions and significance of cyclin D2 and bcl-2 in diffuse large B-cell lymphoma (DLBCL) and their clinical significances.Methods:The tissues of 87 DLBCL patients undergoing resection and 23 patients with lymphoid tissue reactive hyperplasia (RLH) in the First Central Hospital of Baoding from January 2015 to March 2020 were collected. Immunohistochemistry method was used to detect the expressions of cyclin D2 and bcl-2 in tissues of DLBCL and RLH, and the relationship between the expressions of cyclin D2 and bcl-2 as well as the association with the clinicopathological features of DLBCL patients.Results:The positive rates of cyclin D2 protein in DLBCL and RLH were 33.3% (29/87) and 2.0% (1/23), the positive rates of bcl-2 protein in DLBCL and RLH were 60.9% (54/87) and 7.0% (3/23); the positive rates of cyclin D2 and bcl-2 in DLBCL were higher than those in RLH, and the differences were statistically significant( χ2=7.566, P=0.006; χ2=17.512, P < 0.01). The expressions of cyclin D2 and bcl-2 proteins were related to the Ann Arbor staging and immunophenotype of DLBCL patients (all P < 0.05), while not related to age, gender, cancer location, tissue type (all P > 0.05). There was a positive correlation between the expressions of cyclin D2 and bcl-2 protein in DLBCL ( r=1.000, P < 0.01). Conclusions:cyclin D2 and bcl-2 may be related to the development and progression of DLBCL, and both may have some synergies.
ABSTRACT
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.
Subject(s)
Polymorphism, GeneticABSTRACT
Objective According to effect of Sorafenib combined Bevacizumab in mice U14 cervical cancer cell lines,observe the changes of cell lines of transplanted tumor U14 cell structure and adjustment effect of Bcl-2 and Bax protein expression.Methods Sixty female 615 mice of healthy inbreeding line,aged from 4 to 6 weeks,average weight is 20.4 g,range from 18 to 25 g.The animals were divided into blank control group (group A)with 10 mice,and U14 cervical cancer cell lines by vaccinating mice a tumor-burdened mice model was established,on the 6th day after subcutaneous inoculation of U14,40 tumor-burdened mice with tumor diameter ≥ 5 mm were selected.The mice were divided into 5 groups according to the completely randomized grouping method:model group (group B),Sorafenib group (group C),low-dose Bevacizumab combined with Sorafenib group (group D),the middle dose Bevacizumab combined with Sorafenib group (group E),and the high dose Bevacizumab combined with Sorafenib group (group F),8 mice in each group.The treatment regimen consisted of 1 ml of 0.9％ sodium chloride solution in group A and group B,Sorafenib 12 mg/kg in group C,and Bevacizumab 2.5 mg/kg + Sorafenib 12 mg/kg in group D,Bevacizumab 5 mg/kg + Sorafenib 12 mg/kg in group E,and Bevacizumab 10 mg/ kg + Sorafenib 12 mg/kg in group F.All mice were injected intraperitoneally and sacrificed by dislocation 24 days later.The tumor weight (g) was measured every 6 days,and the body weight of each group was observed,and calculated the tumor inhibition rate of each group of mice.The histopathological changes of the transplanted mice were observed by hematoxylin-eosin staining.The transplantation of each group of mice was observed by electron microscope.Morphological changes of tumor tissue;the expression of Bcl-2 and Bax protein in each group were observed by immunohistochemistry.The measurement data were expressed as mean standard deviation (Mean ± SD),univariate analysis of variance was used for inter-group comparison,and LSD method was used for pairwise comparison.Results At the beginning of the intraperitoneal injection of the drug,the body weight of the mice began to increase slowly and continuously,and the trend of the mice in each group was basically the same.The tumor inhibition rate of group C was 8.02％,4.92％ in group D,11.10％ in group E,and 5.42％ in group F.Group E had the highest tumor inhibition rate and the best effect.The difference in tumor weight between group A and the other groups was statistically significant (P ＜ 0.05).The results of electron microscopy showed that the cell structure changes in C,D,E and F groups were similar,and the apoptosis of tumor cells appeared after treatment.The apoptosis of group E was the best.The apoptosis-related proteins in group C,D,E and F were observed.The expression of Bax was up-regulated,and the positive number in group E was the highest.The expression of proto-oncogene Bcl-2 was down-regulated in group C,D,E and F,and the number of positive in group E was the least,and the difference was statistically significant (P ＜ 0.05).Conclusions Sorafenib combined with Bevacizumab can promote tumor cell apoptosis by up-regulating Bax and down-regulating Bcl-2 protein expression.Among them,Sorafenib combined with Bevacizumab at medium dose promotes tumor cell apoptosis better than other methods,providing a basis for clinical treatment.
ABSTRACT
Objective@#To investigate the association between the promoter region-938 polymorphism of B-cell lymphoma/leukemia-2 (Bcl-2) gene and the esophageal cancer (EC) and gastric cardia adenocarcinoma (GCA) in Hebei Province. @*Methods@#From 2007 to 2010, 145 esophageal cancer patients and 169 cardiaccancer patientsfrom the outpatient department of the Fourth Hospital of Hebei Medical Universitywereselected in a case group, and 195 non-tumor patients were selected in a control group during the same period. A questionnaire survey was used to collect information of research subjects. Pathological tissues were collected to extract genomic DNA and detect the genotype of bcl-2 gene -938. A multivariate logistic regression model was used to analyze the association between the bcl-2 gene locus 938 CC genotype and the EC and GCA. The interaction between age, gender, smoking, drinking, upper gastrointestinal family history and the bcl-2 gene locus 938 CC genotype was analyzed by likelihood ratio test. @*Results@#The age of the esophageal and cardiac cancer groups was (56.3±8.3) and (57.1±8.4) years old, and that of the control group was (54.7±7.1) years old. The proportion of the bcl-2 gene locus 938 CC genotype in the esophageal group [48.3% (70/145)] and the cardiac cancer group [48.5% (82/169)] was higher than that in the control group [33.8% (66/195)] (both P values<0.05).Compared with the AA genotype, the risk of esophageal cancer and cardiac cancerin people with the CC genotype was 2.386 (1.20-4.76) and 2.564 (1.27-5.18) respectively. In the population with CC genotype, compared with the positive family history, drinking, and male, the negative family history, non-drinking, and female had a higher risk of esophageal cancer; compared with the non-smoking, negative family history, non-drinking and male, the smoking, positive family history, drinking, and female had a higher risk of cardiac cancer (all the P interaction values were <0.05). @*Conclusion@#People with bcl-2 gene locus 938 CC genotype in Hebei Provincewere more likely to suffer from the esophageal and gastric cardia adenocarcinoma.
ABSTRACT
Objective To investigate the effects of rapamycin on the biological behaviors of human non-Hodgkin lymphoma Raji cells with different concentrations and time, and to explore its mechanism. Methods Different concentrations (0, 10, 50, 100, 250, 500 nmol/L) of rapamycin were used in Raji cells for 24, 48, 72 h respectively. The apoptosis of Raji cells was detected by using CCK-8 method, and flow cytometry was used to detect the cell apoptosis and cycle of Raji cells. The enzymatic activity of Caspase-3 and Caspase-9 in Raji cells was detected by Caspase-3 and Caspase-9 activity testing kit. The expressions of bcl-2, p53 protein and mRNA were detected by Western blot method and reverse transcription-polymerase chain reaction (RT-PCR). Results The proliferative inhibition rate of Raji cells was increased from (23.7 ± 4.2)％to (51.7±3.7)％, the cell apoptosis rate was increased from (4.9±1.9)％to (20.5±1.5)％, the proportion of G0/G1 was increased from (40.8±1.4) ％to (63.6±1.7) ％, the Caspase-3 enzyme activity of Raji cell in 24 h was increased from 0.16±0.05 to 1.08±0.04, Caspase-9 enzyme activity was increased from 0.19±0.04 to 1.34± 0.06 after 24 h with the increasing concentration of rapamycin from 0 nmol/L to 500 nmol/L (P<0.01). The mRNA of bcl-2 was decreased from 0.90±0.03 to 0.46±0.03, and mRNA of p53 was increased from 2.51±0.41 to 5.85±0.21. The protein expression of bcl-2 was reduced and the protein expression of p53 was increased. The experimental results of Raji cells in 48 h and 72 h were consistent with the experimental results of 24 h. Conclusion Rapamycin may inhibit the proliferation of Raji cells through Caspase-3, Caspase-9, bcl-2, p53 and induce its cell apoptosis.
ABSTRACT
Objective To observe the effects of the Diaomo-Zhibeng liquid on the expression of Bcl-2/Bax mRNA and their protein in vitro endometrial glandular epithelial cells in patients of endometrial hyperplasia Methods The endometrial tissue in patients with the pathological diagnosis of endometrial hyperplasia were collected.After isolation,culture and identification,the endometrial cells were divided into four groups:low dosage group (12.5 μg/ml),middle dosage group (25 μg/ml),high dosage group (50μg/ml) and blank control group at randomaccording to the random number table.Three duplicate samples were set for each group.After 48h,the expression of Bcl-2 and Bax were detected by RT-PCR and Western-blot.Results Compared with the blank control group,the expression of Bcl-2 mRNA (1.51 ± 0.07,1.09 ± 0.06,0.93 ± 0.07 vs.1.92 ± 0.08) and protein (0.91 ± 0.02,0.72 ± 0.03,0.62 ± 0.03 vs.1.28 ± 0.02) significantly decreased,while the expression ofBax mRNA (5.73 ± 0.37,8.00 ± 0.23,10.72 ± 0.40 vs.3.27 ± 0.34) and protein (0.45 ± 0.02,0.63 ± 0.02,0.78 ± 0.03 vs.0.32 ± 0.02) sifnificantly promoted in low dosage group,medium dosage group and high dosage group of Diaomo-Zhibeng liquid (P＜0.05).Compared with low dosage group,the expression of Bcl-2 mRNA and protein significantly decreased in middle and high dose group (P＜0.05),but Bax mRNA and protein significantly increased (P＜0.05).Conclusions The Diaomo-Zhibeng liquid can increase the susceptibility of epithelial cells to apoptosis,regulate the balance of cell proliferation and apoptosis tends to apoptosis,have the effect of accelerating cells apoptosis.
ABSTRACT
Objective To investigate the effects of artesunate combined with arsenic trioxide (ATO) on the proliferation and apoptosis of NB4 cells.Methods The NB4 cells were treated with different concentrations of artesunate and arsenic trioxide respectively for 48 h.The cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method.The cells were divided into 4 groups:control group,artesunate group,arsenic trioxide group,and the combination of artesunate and arsenic trioxide group.The cell cycle and apoptosis were detected by flow cytometry (FCM).The protein expression levels of Bcl-2 and Bax were detected by Western blot.Results The MTT results showed that compared with the control group,the proliferation inhibition rates of 0.25,0.50,1.00,2.00,4.00 μmol/L artesunate group (19.26％ ± 3.59％,36.53％ ± 2.67％,61.32％ ± 2.50％,70.30％ ± 3.15％,86.92 ± 0.02％) significantly increased (P＜0.05);the proliferation inhibition rates of 1,2,4,8,16 μmol/L arsenic trioxide group (12.69％ ± 2.43％,64.26％ ± 2.02％,85.10％ ± 2.67％,92.06％ ± 2.21％,93.67％ ± 3.36％) significantly increased (P＜0.05);and the proliferation inhibition rate (40.17％ ± 5.49％ vs.32.23％ ± 3.52％) of combination of artesunate and arsenic trioxide group significantly higher than the arsenic trioxide group (P＜0.05).Compared with the arsenic trioxide group,the percentage of G0/G1 phase cells (74.20％ ± 1.43％ vs.66.14％ ± 1.78％),the apoptosis rate (58.00％ ± 2.41％ vs.34.57％ ± 1.22％),and the expression level of Bax protein (1.35 ± 0.09 vs.1.13 ± 0.09) in the combination of artesunate and arsenic trioxide group significantly increased (P＜0.05),the expression level of Bcl-2 protein (0.45 ± 0.09 vs.1.03 ± 0.10) in the combination of artesunate and arsenic trioxide group significantly decreased (P＜0.05).Conclusions Artesunate can significantly enhance the proliferation inhibition and apoptosis induced by arsenic trioxide on NB4 cells.The possible mechanism of proliferation inhibition and apoptosis of NB4 cells by artesunate combined with arsenic trioxide may be related to reduce the expression of anti-apoptotic protein Bcl-2 and increase the expression of apoptotic protein Bax.
ABSTRACT
Objective To investigate the relationship between single nucleotide polymorphisms of Bcl-2 related anti apoptotic protein 3 (BAG3) gene and Keshan disease (KD) in north Chinese Han population.Methods In 2002 a total of 285 Chinese Han subjects,including 79 KD patients and 206 control subjects were involved in this study.Genomic DNA was extracted from the peripheral venous blood sample.Blood samples were provided by the Institute of Endemic Disease Prevention,Xi'an Jiaotong University,and stored at 80 ℃.The polymorphism of genetic variation was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF).The data was analyzed using TYPER 4.0 or SPSS16.0 software.All sample groups were tested for Hardy-Weinberg equilibrium using goodness-of-fit x2 test.Differences in genotype distribution and allele frequencies between case and control were compared by x2 test.Logistic regression analysis was applied to detect association using age as a confounding factor.Results All sample group passed the Hardy-Weinberg equilibrium test (P ＞ 0.05).Significant differences were not observed in genotype distribution between cases (rs2234962:CC,CT,TT were 0.0％,0.0％ and 100.0％,respectively;rs196295:GG,GA,AA were 22.8％,54.4％ and 22.8％,respectively;rs3858339:GG,GT,TT were 5.1％,38.0％ and 56.9％,respectively;rs3858340:TT,TC,CC were 5.1％,38.0％ and 56.9％,respectively) and controls (rs2234962:CC,CT,TT were 0.0％,1.0％ and 99.0％,respectively;rs196295:GG,GA,AA were 21.4％,51.5％ and 26.2％,respectively;rs3858339:GG,GT,TT were 5.8％,34.5％ and 59.7％,respectively;rs3858340:TT,TC,CC were 5.8％,34.5％ and 59.7％,respectively) for rs2234962,rs3858339,rs196295 and rs3858340 on BAG3 gene (x2 =0.685,0.408,0.330,0.330,P ＞ 0.05).Significant differences were not observed in genotype after agecorrecting between cases and controls for 4 SNPs on BAG3 gene (x2 =0.001,0.019,1.009,0.019,P ＞ 0.05).Conclusion The results suggest that the BAG3 gene might not be a susceptibility gene of KD in north Chinese Han population.
ABSTRACT
Objective To investigate the effect of the over-expressed CTCF on apoptosis factors Bax and Bcl-2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF,Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDA-MB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl-2 were 4.59 ± 0.97 and 10.68 ± 1.93, and there was significant difference between the two groups (t=21.72, P<0.05). Conclusion The over-expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.
ABSTRACT
Objective To explore the research of Jianpi-Huatan decoction resisting the nude mouse MFC hepatic metastasis and its mechanism. Methods MFC cells inoculation in nude mice spleen, establishing nude mice hepatic metastasis model, which are divided into model group, 5-fu injection group, Jianpi-Huatan decoction high, medium and low dose group according to radom number table. Mice in high, medium and low dose Jianpi-Huatan decoction groups were adiminstrated with 80,40 and 20 g/kg Jianpi-Huatan decoction,in 5-fu groups were adiminstrated by intraperitoneal injection with 60mg/kg 5-fu injection and in model groups with Physiological saline once a day for 4 consecutive weeks. After the end, calculate the nude mice weight, spleen tumor weight and evaluation of hepatic metastases. And immune histochemical method and RT-PCR method is applied to detect tumor tissue of the expression of P53, Bcl-2 protein and mRNA. Results Compared with model group, Jianpi-Huatan decoction high and medium dose group can obviously increase body weight[(21.40 ± 1.43)g, (21.70 ± 1.02)g vs.(20.37 ± 1.17)g] and reduce the in situ tumor weight [(0.26 ±0.13)g,(0.16 ±0.05)g vs.(0.63 ±0.17)g]and the number of liver metastases;the mRNA levels of P53 and Bcl-2 (8.32 ±0.38,5.42 ±0.45,3.09 ±0.26 vs.9.67 ±1.31)and(4.65 ±0.61,3.22 ±0.21,2.49 ±0.19 vs.5.32 ±0.42) were decreased in low,medium and high dose Jianpi-Huatan decoction groups;P53(76.11 ±5.23,45.20 ±3.77, 23.11 ± 3.14 vs.81.63 ± 5.01)and Bcl-2(58.67 ± 5.27,32.00 ± 3.13,19.00 ± 2.54 vs.63.00 ± 4.10)levels were down-regulated in each dose of Jianpi-Huatan decoction group.Conclusions Jianpi-Huatan decoction can restrain nude mouse transplantation tumor growth and hepatic metastasis, which related to the cut of the expression of P53, Bcl-2 gene and protein.
ABSTRACT
Objective To investigate the effect of the over-expressed CTCF on apoptosis factors Bax and Bcl-2 in human breast cancer cell line MDA-MB-231. Methods Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expressions of CTCF,Bax and Bcl-2 in MDA-MB-231. The overexpression vector of CTCF/pEGFP-N1 was constructed. The overexpression plasmid CTCF/pEGFP-N1 and the empty vector plasmid pEGFP-N1 were transfected into breast cancer cell line MDA-MB-231 by lentivirus transfection, and the MDA-MB-231 cells were divided into CTCF group and control group. After successfully transfection of MDA-MB-231 identified by RT-PCR, real time quantitative PCR (Q-PCR) was used to detect the mRNA levels of Bax and Bcl-2 in MDA-MB-231 of the CTCF group and the control group. The protein levels of Bax and Bcl-2 were detected by Western blot assay and enzyme-linked immunosorbent assay (ELISA). Results The expression of CTCF was not found in MDA-MB-231, and expressions of Bax and Bcl-2 were found in MDA-MB-231. Results of Q-PCR showed that the mRNA levels of Bax were 4.63±1.08 and 2.27±0.16 in CTCF group and control group, respectively, and they were statistically significant (t=27.50, P<0.05). The mRNA levels of Bcl-2 were 1.39±0.14 and 3.56 ± 0.97 in CTCF group and control group, and there was significant difference between two groups(t=39.00, P<0.05). Results of Western blot assay showed that the protein level of Bax was higher in CTCF group compared with that of control group. The protein level of Bcl-2 was lower in CTCF group compared with that of control group. Results of ELISA showed that the protein levels of Bax were 15.25±2.17 and 6.24±1.78 in CTCF group and control group, respectively, and there was significant difference between the two groups (t=26.84, P<0.05). The protein levels of Bcl-2 were 4.59 ± 0.97 and 10.68 ± 1.93, and there was significant difference between the two groups (t=21.72, P<0.05). Conclusion The over-expressed CTCF can promote the expression of apoptotic factors and inhibit the expression of anti-apoptotic factors in breast cancer cells.
ABSTRACT
Objective To detect the expression of bcl-2 and c-Met genes in lung cancer cell lines with different mutations of epidermal growth factor receptor (EGFR), in order to explore the association between expression of bcl-2 and c-Met genes and drug resistance in non-small cell lung cancer (NSCLC). Methods Direct sequencing was used to detect EGFR mutations status in HCC827 cells, A549 cells and H1975 cells. Immunocytochemistry was conducted to test bcl-2 and c-Met expression. RT-PCR was performed to analyzed bcl-2 gene expression and ARMS was used to detect EGFR mutations status in malignant pleural effusion of NSCLC patients. Results A549 cells, HCC827 cells and H1975 cells were EGFR wild type, EGFR exon 19 deletion (19del), and EGFR exon 21 L858R and exon 20 T790M double mutations. c-Met and bcl-2 protein located in cytoplasm and the intensity of positive expression was highest in HCC827 cells, followed by A549 cells and H1975 cells. The bcl-2 mRNA expression was higher in HCC827 and A549 cells than that in H1975 cells (10.93±1.90 vs. 0.83±0.15, P=0.013; 7.13±1.33 vs. 0.83±0.15, P= 0.000). However bcl-2 mRNA expression was not associated with EGFR mutations (wild type, 19del and L858R) in malignant pleural effusion of NSCLC patients. Conclusion bcl-2 and c-Met gene in HCC827 cells (EGFR 19del) expression is significantly higher than those in H1975 cells (EGFR L858R/T790M), implying EGFR L858R mutations and 19del mutations may be regulated by different signaling pathways.
ABSTRACT
PURPOSE: T o investigate the possible protective effect of thymoquinone (TQ) in cisplatin (CP) induced myocardial injury. METHODS: A total of 28 adult male Wistar-Albino rats were randomly and equally divided into four groups as follows: Group 1 (control), Group 2 (CP at 15 mg/kg dose), Group 3 (TQ 40 mg/kg/day for two days prior to CP injection and on third day, CP at 15 mg/kg dose was intraperitoneally administered and TQ treatment continued until fifth day) and Group 4 (TQ at 40mg/kg/day dose for five days). RESULTS: There was a significant increment in CP group in terms of congestion, edema and pycnotic nuclei in myocardial fibers, comparing with other groups. TQ group exhibited significant increase in expression of antiapoptotic protein Bcl-2, comparing with CP group (p<0.05). In only CP administered group, expression of antiapoptotic protein Bcl-2 was lowest comparing with other groups. CONCLUSION: Established data indicate that cisplatin is cardiotoxic and thymoquinone may be useful in treating CP-induced cardiac injury.
Subject(s)
Animals , Male , Benzoquinones/pharmacology , Cisplatin/toxicity , Cardiomyopathies/chemically induced , Cardiomyopathies/prevention & control , Antineoplastic Agents/toxicity , Antioxidants/pharmacology , Reference Values , Time Factors , Immunohistochemistry , Random Allocation , Reproducibility of Results , Benzoquinones/therapeutic use , Treatment Outcome , Rats, Wistar , Apoptosis/drug effects , Oxidative Stress/drug effects , Proto-Oncogene Proteins c-bcl-2/analysis , Proto-Oncogene Proteins c-bcl-2/drug effects , Myocytes, Cardiac/drug effects , Cardiotoxicity/etiology , Cardiotoxicity/pathology , Cardiotoxicity/prevention & control , Heart/drug effects , Cardiomyopathies/pathology , Myocardium/pathology , Antioxidants/therapeutic useABSTRACT
The B-cell lymphoma gene 2 (Bcl-2) is considered as the most important inhibiting cell apoptosis control gene,and its expression is up-regulated in asthma.Drug intervention against Bcl-2 affects signal pathways,down-regulates the Bcl-2 expression,and induces programmed cell death.It may become the target of asthma treatment.This article reviews the structure characteristics of the Bcl-2,the mechanism of regulating apoptosis,and its role in asthma.
ABSTRACT
Objective To investigate the effects of fasudil on expressions of Bcl-2 and Bax in cerebral cortex of model rats with subarachnoid hemorrhage (SAH). Methods Thirty rats were divided into sham operation group, SAH group and SAH+fasudil group, 10 rats in each group. Double injection of blood into occipital cistern method was used for SAH model in SAH group and SAH+fasudil group. In the sham operation group, the blood injection was instead by normal saline. In the SAH+fasudil group, at 30 min after the second injection of blood, rats were administrated with intraperitoneal injection of fasudil at a dose of 3 mg/kg. The general situation, neurological score, TUNEL staining for cortex cell apoptosis, immune histochemical staining and Western blotting assay for Bcl-2 and Bax protein expression were compared 24 h after the operation between the three groups. Results Compared with the sham operation group, rats in SAH group and SAH +fasudil group appeared obvious neurological deficits. The neurological score was significantly lower in SAH group ( 2.68 ± 0.31) than that of sham operation group (5.00±0.00). The neurological score was significantly higher in SAH+fasudil group (3.27 ± 0.35) compared with that of SAH group (2.68 ± 0.31, P<0.05). There was obvious cell apoptosis in SAH group and SAH+fasudil group, and the apoptosis was less in SAH+fasudil group than that of SAH group (P<0.05). The level of Bcl-2 expression was significantly lower in SAH group than that of sham operation group, and Bax expression was significantly higher in SAH group than that of sham operation group (P<0.05). The level of Bcl-2 expression was significantly higher in SAH+fasudil group than that of SAH group, but Bax expression was significantly lower in SAH+fasudil group than that of SAH group (P<0.05). Conclusion Fasudil can improve the neurological damage in rats with SAH, which may be related with the regulation of apoptosis related proteins Bcl-2 and Bax.
ABSTRACT
Objective To observe the effect of the active ingredient present study curcumol impact on human glioma cell line U251 proliferation and apoptosis,its effect on the detection of apoptosis-related gene expression in glioma,and explore the possible mechanism of its anti-glioma,Chinese herbal medicine for the treatment of glioma accumulation of experimental data.Methods In cultured human glioma cell line U251 as a model to medium dilution Curcumol different concentrations,observation and detection following:MTT assay at different times,different concentrations curcumol on U251 cell proliferation;microscope Effect of different concentrations of curcumol U251 cell morphology;affected by flow cytometry curcumol different concentrations on apoptosis in U251;RT-PCR method to detect different concentrations curcumol on U251 cell apoptosis-related gene bcl-2 and COX-2 expression levels of influence.Results Curcumol U251 human glioma cell proliferation inhibition effect and showed concentration-dependent and time-dependent manner (P ＜ 0.05).Conclusions Curcumol U251 human glioma cell proliferation clear.Induce apoptosis and inhibit proliferation of glioma should be an important mechanism of inhibition of proliferation effect of curcumol.Curcumol down on U251 cells bcl-2 gene expression levels and COX-2should be its induction of apoptosis,an important mechanism of inhibition of proliferation.
ABSTRACT
Objective To investigate the effect of highly expressed β-catenin on the proliferative activity of and expressions of two apoptosis-related genes Bcl-2 and Bax by human skin fibroblasts induced by hydrogen peroxide (H2O2).Methods Normal human skin fibroblasts (HSFs) from child foreskin were divided into three groups:empty vector group transfected with the empty vector pcDNA3.1,H2O2 group transfected with the empty vector pcDNA3.1 followed by treatment with H2O2 (150 μ mol/L) for 2 hours,β-catenin group transfected with pcDNA3.1-β-catenin followed by treatment with H2O2 (150 μ mol/L) for 2 hours.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was conducted to estimate proliferative activity of fibroblasts,flow cytometry to detect cell apoptosis,and reverse transcription (RT)-PCR and Western blot were performed to measure the mRNA and protein expressions of Bcl-2 and Bax respectively.The relative expression levels of genes were expressed as the ratios between the targets and GAPDH.Results Significant differences were found between the empty vector group,H2O2 group and β-catenin group in cellular proliferative activity (expressed as absorbance value at 570 nm:0.792 ± 0.012 vs.0.462 ± 0.012 vs.0.521 ± 0.015,P＜ 0.01) and apoptosis rate (3.407％ ± 0.217％ vs.24.555％ ± 1.793％ vs.15.360％ ± 0.755％,P＜ 0.01).Both mRNA and protein expression levels of Bcl-2 were significantly lower in the H2O2 group (0.333 ± 0.003 and 0.336 ± 0.004 respectively) than in the empty vector group (0.507 ± 0.013 and 0.514 ± 0.021,respectively,both P ＜ 0.01) and β-catenin group (0.404 ± 0.006 and 0.411 ± 0.005,respectively,both P ＜ 0.01).Increased expression levels of Bax mRNA and protein were observed in the H2O2 group compared with the empty vector group and β-catenin group (mRNA:0.451 ± 0.002 vs.0.303 ± 0.005 and 0.339 ± 0.012,protein:0.460 ± 0.008 vs.0.320 ± 0.013 and 0.346 ± 0.013,all P＜ 0.01).Conclusion High expression of β-catenin can raise proliferative activity of aging HSFs.
ABSTRACT
Objective To explore the inhibitory effects of lobaplatin and cisplatin and their regulation of apoptosis-related genes in ovarian cancer cells in nude mice.Methods SKOV3 cells were implanted into nude mice.In monotherapy treatment study,the nude mice bearing human SKOV3 cells were randomly divided into control,lobaplatin,and cisplatin groups,with 7 mice in each group.The mice in each group were received corresponding treatment.The volume of tumor and the weight of nude mice were measured three times per week,respectively.Tumor inhibitory rate was calculated.The protein expressions of bax and bcl-2 were detected by flow cytometry.Results The growth inhibitory rate was 47.2％ in lobaplatin group and 42.8％ in cisplatin group,without significant difference between two groups (P ＞ 0.05).The expression of bcl-2 was decreased but the bax was increased in lobaplatin and ciaplatin groups compared to the control group.Conclusions Lobaplatin can significantly inhibit the growth of ovarian cancer cells,induce apoptosis by down-regulation of bcl-2 and up-regulation of bax.
ABSTRACT
Objective To inveatigate the expression and clinical significance of microRNA‐204(miR‐204) in human renal clear cell carcinoma(RCCC) .Methods 65 cases of resected RCCC tissue and corresponding normal tumor‐adjacent tissue were col‐lected and detected the expression level of miR‐204 by using qRT‐PCR .The correlation between the miR‐204 expression level and clilicalpathological features was statistically analyzed .The artificial miR‐204 mimics were adopted to stranfect into Caki‐1 cells ,then the Western blot was used to detect the expression of Bcl‐2 a as the downstream potential targets of miR‐204 and SIRT1 mRNA and protein .Results The expression level of miR‐204 in the RCCC tissue was significantly lowere than that in the normal tumor‐adjacent tissues (P3 cm ,P<0 .05) , lymph node metastasis(P<0 .05) and advanced TNM stage(Ⅲ + Ⅳ ,P<0 .05) .Both the mRNA and protin expression in RCCC Caki‐1 cells were down‐regulated after transfection .Conclusion The expression of miR‐204 is down-regulated in the RCCC tissue and is related to the malignant clinicopathological features ,and miR‐204 may suppress the RCCC genesis and development through down‐regulating the expression of Bcl‐2 and SIRT1 .