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Objective:To explore the effects of BCR-ABL downstream pathway inhibitors, such as RAF inhibitor SB590885, JAK inhibitor AZD1480, PI3K-mTOR double target inhibitor BEZ235 on chronic myelogenous leukemia (CML) cells, and the effect of BEZ235 on the proliferation, apoptosis of CML cells and the sensitivity of imatinib in vitro.Methods:K562 cells were treated with different concentrations of the drugs. MTS method was applied to detect the proliferation inhibition rate of K562 cells, and 50% inhibitory concentration (IC 50) of all drugs for 48 h was calculated. The cell apoptosis rate was tested by using flow cytometry with Annexin V-FITC/PI double staining. The cell cycle was tested by using flow cytometry with PI staining. K562 cells, imatinib-resistant and T315I-mutant human CML KBM7R cells and imatinib-resistant CML primary cells of patients were treated with different concentrations of the drugs. MTS method was used to test the proliferation inhibition of cells, and IC 50 of all drugs for 48 h was evaluated. KBM7R cells or primary cells of CML patients were treated with 1.0 μmol/L BEZ235, 1.0 μmol/L imatinib or the combination of both, respectively. Flow cytometry with PI staining was used to detect the cell cycle of KBM7R cells. Flow cytometry with Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate in CML primary cells. The expressions of p-AKT, cleaved Caspase-3 and Cyclin D1 proteins were detected by using Western blot. Results:SB590885, AZD1480 and BEZ235 could inhibit the proliferation of K562 cells, and the IC 50 after the treatment of K562 cells for 48 h was (11.49±3.14), (4.83±1.26) and (0.37±0.21) μmol/L, respectively. SB590885, AZD1480 and BEZ235 could promote the apoptosis of K562 cells. The cell apoptosis rates were increased compared with the control group without drug treatment (all P < 0.01). SB590885 and BEZ235 induced G 0/G 1 block (both P < 0.05). AZD1480 induced G 2/M block ( P < 0.05). BEZ235 could inhibit the proliferation of K562 cells, KBM7R cells and CML primary cells, and their IC 50 for 48 h was (0.37±0.21), (0.43±0.27) and (0.49±0.24) μmol/L, respectively. Compared with imatinib alone, the different concentrations of imatinib combined with 0.2 μmol/L BEZ235 could increase the proliferation inhibition of K562 cells, KBM7R cells and CML primary cells, and could reduce the IC 50 of imatinib. After the treatment of imatinib alone and combination with BEZ235 for 48 h, the imatinib IC 50 of K562 cells was (0.14±0.05) and (0.09± 0.04) μmol/L ( t = 1.531, P = 0.249), the imatinib IC 50 of KBM7R cells was (3.93±2.29) and (0.44±0.22) μmol/L ( t = 2.837, P = 0.047), the imatinib IC 50 of the primary cells was (3.12±1.53) and (0.39±0.23) μmol/L ( t = 3.925, P = 0.042). The cell apoptotic rate of the primary cells was (4.9±1.4)%, (13.1±3.2)%, (8.8±2.0)% and (40.6±6.0)%, respectively in the control group without drug treatment, 1.0 μmol/L BEZ235, 1.0 μmol/L imatinib and the combination of 1.0 μmol/L BEZ235 and 1.0 μmol/L imatinib after the treatment of 24 h ( F = 71.031, P < 0.01). Compared with imatinib alone, the expressions of p-AKT and Cyclin D1 proteins were decreased, and the expression of cleaved Caspase-3 protein was increased after the treatment of KBM7R cells for 12 h in the combination group of both BEZ235 and imatinib. Conclusions:BCR-ABL downstream pathway inhibitors can effectively inhibit the proliferation and promote the apoptosis of K562 cells. BEZ235 can inhibit the proliferation and promote the apoptosis of K562 cells, imatinib-resistant and T315I-mutant human KBM7R cells and imatinib-resistant CML primary cells of patients, which has a synergistic effect to imatinib.
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The over-activation of phosphatidylinositol 3-kinase (PI3K) signaling pathway is closely related to the occurrence and development of malignant tumors. The abnormal expression of this pathway is involved in the regulation of cell growth, malignant transformation, apoptosis and metastasis. The targeted therapy for specific sites of PI3K signaling pathway is a new research hotspot. A variety of different types of PI3K inhibitors have been marketed or entered clinical trials and have shown considerable efficacy in the treatment of malignant lymphomas. In this view, the recent advances in activation patterns of PI3K signaling pathway in lymphoma and PI3K inhibitors are summarized.
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The over-activation of phosphatidylinositol 3-kinase (PI3K) signaling pathway is closely related to the occurrence and development of malignant tumors. The abnormal expression of this pathway is involved in the regulation of cell growth, malignant transformation, apoptosis and metastasis. The targeted therapy for specific sites of PI3K signaling pathway is a new research hotspot. A variety of different types of PI3K inhibitors have been marketed or entered clinical trials and have shown considerable efficacy in the treatment of malignant lymphomas. In this view, the recent advances in activation patterns of PI3K signaling pathway in lymphoma and PI3K inhibitors are summarized.
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Objective To assess the activation of PI3K-AKT-mTOR signal pathway in Burkitt lymphoma.Methods The phosphorylation levels of AKT,mTOR and RPS6 were detected in the tissue sections by immunohistochemistry in 13 cases of Burkitt lymphomas and 14 cases of reactive hyperplasia of lymph nodes.Results The phosphorylation rates of AKT,mTOR and RPS6 were higher in Burkitt lymphoma than those in reactive hyperplasia of lymph nodes [84.6 ％(11/13) vs 64.2 ％(9/14),100.0 ％(13/13) vs 71.4 ％ (10/14),100.0 ％ (13/13) vs 78.6 ％ (11/14),all P ＜0.05].Conclusion PI3K-AKT-mTOR pathway is abnormally activated in Burkitt lymphoma.
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Objective The goal of the study was to assess the lethal effect of cytotoxic lymphocyte against A549 cells induced by dendritic cells (DC) pulsed with A549 lysate and transfected with GM-CSF recombinant adenovirus. Methods The cytotoxic lymphocyte against A549 cells were induced by culturing with DCs, which pulsed with A549 antigens and transfected with GM-CSF recombinant adenovirus. The of effector/target ratio, the killing rates of N-DC group, A-DC group and G-A-DC group were (1.9±0.7) %,effector/target ratio, the killing rates of N-DC group, A-DC group and G-A-DC group were (5.3±0.2) %, (40.5±7.7) % and (72.5±4.7) %, respectively. We found that the killing rate of G-A-DC group was the highest by statistics. Conclusion The cytotoxic lymphocyte against A549 cells can be induced by DCs pulsed with A549 lysate ,and the lethal effect of CTLs can be enhanced when DCs were infected with GM-CSF recombinant adenovirus.
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ObjectiveTo investigate the expression of survivin,COX-2 and bcl-2 in non-Hodgkin lymphoma(NHL)and the significance and correlation between them.MethodsImmunohistochemistry MaxVision systems for survivin,COX-2 and bcl-2 were conducted on 44 NHL and 20 reactive lymphoid hyperplasia (RH).ResultsThe positive expression rates of survivin,COX-2,bcl-2 in NHL were 70.45 ％(31/44),68.18 ％ (30/44),63.64 ％ (28/44),respectively,and these in RH were 40.00 ％ (8/20),40.00 ％(8/20) and 20.00 ％ (4/20),respectively.There was positive correlation between the expression of COX-2 and survivin (r =0.306,P =0.043),survivin and bcl-2 protein (r =0.339,P =0.040) in NHL.ConclusionCOX-2,survivin,bcl-2 are highly expressed in NHL.To detect the expression of them has clinical value to diagnosis NHL and to estimate the malignant degree of lymphoma.There are a positive correlations between the expression of COX-2 and survivin protein,and between the expression of survivin and bcl-2 protein,which indicates that they may play a synergistic role in the occurrence and development of NHL.
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Risk factors of hyperamylasemia in 152 diabetic ketoacidosis(DKA)patients who had no acute pancreatitis were examined.Serum levels of hemodiastase,natrium,kalium,chlorine,calcium,phosphonium,creatinine,carbon dioxide combining power,osmotic pressure,blood glucose,blood lipids and uric acid were measured.With hemodiastase and other 14 factors as independent variables,the multiple linear regression analysis was performed and the partial regression coefficient was estimated.Blood glucose,serum natrium,osmotic pressure and triglyceride entered the regression equation,and their partial coefficients were 10.26,10.35,2.21 and 8.00,respectively.The constant quantitv was -2162.06.Compared with amylase-normal group,hyperamylasemia group showed statistically significant difference in blood glucose,serum natrium,osmotic pressure and triglyceride levels.Higher blood glucose and triglyceride might be the primary causes of hyperamylasemia.