Progress of bcl-2 inhibitors in treatment of plasma cell disorders with t(11;14) / 白血病·淋巴瘤

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.

Expressions of cyclin D2 and bcl-2 in diffuse large B-cell lymphoma and their clinical significances / 白血病·淋巴瘤

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.

[Clinicopathological features of de novo CD5-positive diffuse large B-cell lymphoma].

Objective: To investigate the relationship between the protein expression of C-MYC, bcl-2 and bcl-6 and the clinicopathological characteristics in patients with de novo CD5-positive diffuse large B cell lymphoma (CD5(+)DLBCL). Methods: Fifty seven cases of de novo CD5(+)DLBCL were collected at Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from February 2013 to September 2018. The hematoxylin-eosin stained slides were reviewed, and immunohistochemical (IHC) staining and FISH were used to analyze the relationship between C-MYC, bcl-2, bcl-6 expression and the clinicopathologic characteristics of patients. Results: Among these 57 cases, 27 were male and 30 were female. The age of onset was 35-99 years old. The IHC expression rates of C-MYC, bcl-2 and bcl-6 were 50.9% (29/57), 84.2% (48/57), and 75.4% (43/57) respectively; and co-expression rate of C-MYC and bcl-2 proteins was 40.4 (23/57). There was no significant correlation between protein expression and patients' genders, clinical stage, the level of serum LDH,ß2 microglobulin, IPI,B symptoms, bone marrow involvement and central nervous system recurrence (P>0.05). Univariate analysis showed that the median OS of C-MYC negative patients was significantly longer than C-MYC positive patients (P<0.05); and the median OS of patients without double expression was significantly longer than that of patients with positive expression (P<0.05), and bcl-6 positive patients had longer median OS than bcl-6 negative patients (P<0.05). There was no significant correlation between prognosis and bcl-2 protein expression (P>0.05) . Cox multivariate analysis showed C-MYC protein expression was an independent predictor of OS in de novo CD5(+)DLBCL (P<0.05). Conclusions: Bcl-2 protein expression has no effect on the prognosis in de novo CD5(+)DLBCL whereas bcl-6 expression is correlated with good prognosis. C-MYC protein expression could be used as an independent and effective index to predict the prognosis of patients with de novo CD5(+)DLBCL.However, the relationship between protein expression and gene rearrangement of C-MYC, bcl-2 and bcl-6 needs to be further explored.

Effect and mechanism of ethanol extracts of muxiang (Radix Aucklandiae) on gastric ulcers in rats.

OBJECTIVE: To investigate the effect of ethanol extracts of Muxiang (Radix Aucklandiae) (RA) on gastric ulcers in rats and explore the potential mechanisms. METHODS: A model was established by ethanol (0.75 mL/kg). According to body weight, rats were pretreated with RA extracts (2.5 or 5 g/kg). The rats were administered 95% ethanol orally after 1 h. The effects of ethanol were evaluated by measuring the gastric secretion volume, pH, pepsin activity, and ulcer area. Histological analysis and immunohistochemistry were also conducted. Furthermore, the effect of the ethanol extract of RA on transiting activity of the gastrointestinal tract was observed in mice. RESULTS: Intragastric administration of RA extracts protected the gastric mucosa from ethanol-induced gastric ulcers, while reducing submucosal edema and preventing hemorrhagic damage. Moreover, the extracts increased the production of gastric mucus, upregulated Bcl-2, and downregulated Bax expression. Importantly, pretreated rats exhibited no significant change in the gastric secretion volume, gastric juice acidity, or pepsin. Furthermore, pretreatment prominently (P < 0.05) enhanced propulsive movement of the gastrointestinal tract in normal mice and mice with gastrointestinal motility disorders. CONCLUSION: Ethanol extracts of RA ameliorated gastric lesions in the gastric ulcer rat model. The mechanisms of action were related to improvement of gastrointestinal dynamics, maintenance of mucus integrity, and inhibition of apoptosis by downregulating proapoptotic Bax protein and upregulating anti-apoptotic Bcl-2 protein.

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 association of Bcl-2 gene polymorphism with the esophageal cancer and gastric cardia adenocarcinoma in Hebei Province].

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.

Effect of Qiangxin Huoli decoction on rats with adriamycin-induced chronic heart failure.

OBJECTIVE: To investigate the effect of Qiangxin Huoli decoction on rats with chronic heart failure (CHF) induced by adriamycin (ADR), and to investigate the underlying mechanism of this effect. METHODS: Ninety-six healthy Wistar rats were divided into six groups: control, CHF model, CHF treated by Shenfu injection, and three CHF groups treated with Qiangxin Huoli decoction at high, medium, and low doses, respectively. Qiangxin Huoli decoction was administered orally to protect the stomach in the three Qiangxin Huoli decoction groups, while the control group and the CHF model group were administered the same volume of 0.9% physiological saline, and the Shenfu group wereadministered the same volume of Shenfu injection. Ten days later, the CHF model was then induced in all groups except the control group by intraperitoneal injection of ADR at gradient dose intervals. The bodyweights were recorded on days 10, 20, 30, and 40. Hemodynamic indices were recorded, including left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximum increase in left ventricular pressure (+dp/dtmax), maximum decrease in left ventricular pressure (-dp/dtmax), heart rate (HR), and electrocardiogram using an eight-channel physiological recorder with LabChart software monitoring. The plasma brain natriuretic peptide (BNP) concentration was determined by enzyme-linked immunosorbent adsorption. The expressions of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) were detected by immunohistochemical methods. RESULTS: The CHF model group were in poor condition, and the mean bodyweight was significantly decreased compared with the control group. Furthermore, compared with the control group, the CHF groups had significantly decreased LVSP, +dp/dtmax, and -dp/dtmax, and significantly increased LVEDP. The CHF groups also showed significant increases in HR, S-T segment elevation, and plasma BNP levels compared with the control group. Compared with the CHF model group, the treatment groups had significantly increased Bax expression (P < 0.05) and significantly decreased Bcl-2 expression (P < 0.01), indicating less apoptosis. The high dose Qiangxin Huoli decoction group and the Shenfu group showed the most significant improvements. CONCLUSION: In the rat model of CHF, Qiangxin Huoli decoction significantly reduces the abnormal hemodynamics, improves cardiac function, reduces plasma BNP concentration, regulates the expression of apoptosis proteins, inhibits the apoptosis of myocardial cells, and plays a protective role.

The association of Bcl-2 gene polymorphism with the esophageal cancer and gastric cardia adenocarcinoma in Hebei Province / 中华预防医学杂志

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.

Study on the mechanism of Sorafenib combined with Bevacizumab on the tumor inhibition of cervical cancer cell line U14 transplantation in mice / 国际外科学杂志

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.

An association between single nucleotide polymorphisms of Bcl-2 related anti apoptotic protein 3 gene and Keshan disease in north Chinese Han population / 中华地方病学杂志

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.

Effects of the Diaomo-Zhibeng liquid on the expression of Bcl-2/Bax mRNA and their protein in vitro human endometrial glandular epithelial cells / 国际中医中药杂志

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.

Effect of combination of artesunate and arsenic trioxide on the proliferation and apoptosis of NB4 cells / 国际中医中药杂志

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.

Effect of rapamycin on the biological behaviors of human non-Hodgkin lymphoma Raji cells and its mechanism / 白血病·淋巴瘤

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.

[The role of FOXO3a-Bim signaling in triptolide induced bladder cancer T24 cells apoptosis].

Objective: To investigate the role of FOXO3a-Bim signaling in triptolide induced bladder cancer T24 cells apoptosis. Methods: T24 cells were used and divided into control group, triptolide group(50 nmol/L), MK2206 group(50 nmol/L triptolide+ 5 µmol/L MK2206), FOXO3a-siRNA group(50 nmol/L triptolide+ 100 nmol/L FOXO3a-siRNA), Bim-siRNA group (50 nmol/L triptolide+ 100 nmol/L Bim-siRNA). MTT assay was used to analyze the cells growth inhibition.Annexin V/PI staining was implemented to detect cell apoptosis rate, the expression of p-Akt, Akt, p-FOXO3a, FOXO3a, Bim, Bax.Cleaved-caspase 3 was analyzed by Western blot. Results: After treatment with triptolide 25, 50, 100, 250 nmol/L, the cell growth inhibition rates at 24 hours(17%±9%, 24%±5%, 43%±8%, 61%±8%), 48 hours (20%±7%, 34%±6%, 56%±7%, 74%±5%) and 72 hours(32%±8%, 41%±7%, 69%±7%, 84%±3%) were significantly higher than control group respectively.The IC(50) at 24, 48, 72 hours were (113±10), (91±8), (68±5) nmol/L; the cell apoptosis rates at 24 hours (10%±4%, 15%±5%, 29%±8%, 46%±8%), 48 hours (16%±5%, 24%±6%, 40%±7%, 55%±9%) and 72 hours (27%±4%, 38%±5%, 50%±9%, 65%±8%) were significantly increased (P<0.05). Western blot showed that triptolide reduced the expression of p-Akt, p-FOXO3a and increased the expression of Bim, Bax, cleaved-caspase 3.The cell inhibition rate in Triptolide group (30%±8%) was significantly higher than that in the control group (P<0.05) and the rates in MK2206 group (54% ±6%), FOXO3a-siRNA group (18%±7%) and Bim-siRNA group (11%±6%) were also higher than the control group.Compared with the triptolide group, the inhibition rate in MK2206 group was significantly increased, but decreased in FOXO3a-siRNA group and Bim-siRNA group(P<0.05). Conclusion: Triptolide induces T24 cells apoptosis through FOXO3a-Bim signaling pathway.

The research of Jianpi-Huatan decoction resisting MFC hepatic metastasis and its mechanism / 国际中医中药杂志

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.

Effects of overexpression of CTCF on apoptosis factors Bax and Bcl-2 in breast cancer cells / 天津医药

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.

Effects of overexpression of CTCF on apoptosis factors Bax and Bcl-2 in breast cancer cells / 天津医药

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.

Expression of bcl-2 and c-Met genes in non-small cell lung cancer cell lines harboring different mutations of epidermal growth factor receptor / 肿瘤研究与临床

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.

Effect of chronic intraperitoneal aminoguanidine on memory and expression of Bcl-2 family genes in diabetic rats.

Long-term hyperglycemia associates with memory defects via hippocampal cells damaging. The aim of the present study was to examine the effect of 1 month of i.p. injections of AG on passive avoidance learning (PAL) and hippocampal apoptosis in rat. Eighty male rats were divided into 10 groups: control, nondiabetics and STZ-induced diabetics treated with AG (50, 100, 200, and 400 mg/kg, i.p.). PAL and the Bcl-2 family gene expressions were determined. Diabetes resulted in memory and Bcl-2 family gene expression deficits. AG (50 and 100 mg/kg) significantly improved the learning and Bcl-2, Bcl-xl, Bax, and Bak impairment in diabetic rats. However, negative effects were indicated by higher doses of the drug (200 and 400 mg/kg). Present study suggests that 1 month of i.p. injections of lower doses of AG, may improve the impaired cognitive tasks in STZ-induced diabetic rats possibly by modulating Bcl-2 family gene expressions.

Effects of thymoquinone against cisplatin-induced cardiac injury in rats

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.