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OBJECTIVE@#To analyze relation of ASXL2 gene mutation with the clinical characteristics, prognosis and C-KIT gene mutation in acute myeloid leukemia (AML) patients with AML1-ETO fusion gene.@*METHODS@#The clinical data of 63 primary AML patients with AML1-ETO fusion gene were collected and retrospectively analyzed. The mutation of ASXL2 gene was directly sequenced by PCR. The clinical characteristics, C-KIT mutation rate and prognosis were compared between the patients with ASXL2 gene mutation (group A) and non-mutation (group B).@*RESULTS@#Among 63 patients, 8 (12.70%) cases of ASXL2 mutation gene was detected. Hemoglobin level in peripheral blood of patients in group A was significantly lower than that in group B (P＜0.01). There was no significant difference in sex, ages proportion of bone marrow blasts, lymph node enlargement, peripheral blood leukocytes count and platelets between the two groups (P＞0.05). The infiltration of central nervous system, liver and spleen was not found in both groups. The expression of CD33 in group A was significantly lower than that in group B (P＜0.05), but the results of other immunophenotype analysis were not significantly different between the two groups (P＞0.05). The remission rate and median survival time were not significantly different between two groups (P＞0.05). The detection rate of C-KIT gene mutation were not significantly different between group A and group B (P＞0.05).@*CONCLUSION@#Among AML patients with AML1-ETO fusion gene, ASXL2 gene mutation accounts for a certain ratio, and the peripheral blood hemoglobin concentration and CD33 expression in these patients are often low. At the same time, ASXL2 gene mutation may not be closely related with C-KIT gene mutation.
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OBJECTIVE@#To study the inhibition effect of siRNA on the expression of Wisp-1 gene in Hca-F of mouse hepatocellular carcinoma cells strain and also its effect on the proliferation, migration and adhesion of hepatocellular carcinoma cells.@*METHODS@#Three expression vectors of siRNA were constructed. Lipo2000 was employed to transfect Hca-F cells and Western blot was used to detect the inhibition effect of siRNA on the expression of Wisp-1 gene. Afterward, CCK8 was adopted to detect the effect of Wisp-1 siRNA on the proliferation of Hca-F cells; Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of Wisp-1 siRNA on the apoptosis of Hca-F cells; Transwell was used to detect the effect of Wisp-1 siRNA on the migration of Hca-F cells. The in vitro cell adhesion kit was used to detect of Wisp-1 siRNA on the change in the components of extracellular matrix to which Hca-F cells adhered. Western blot was used to detect the activation of protein kinase B (AKT)/glycogen synthase kinase-3β pathway and the expression of downstream target protein p53 and matrix metalloproteinases-2.@*RESULTS@#The siRNA showed interference effect on the expression of Wisp-1 gene. Compared with the control group, after being transfected to cells, Wisp-1 siRNA could significantly inhibit the proliferation, migration and adhesion of Hca-F cells and also promote the cell apoptosis, which was related to the down-regulated phosphorylation of AKT and glycogen synthase kinase-3β and the expression of p53 and matrix metalloproteinases-2 (P < 0.05).@*CONCLUSIONS@#The inhibition of Wisp-1 expression can reduce the proliferation, migration and adhesion of mouse hepatocellular carcinoma cells, which is related to the AKT/glycogen synthase kinase-3β pathway. Wisp-1 gene may be the potential target to cure the hepatocellular carcinoma.
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Objective: To study the inhibition effect of siRNA on the expression of Wisp-1 gene in Hca-F of mouse hepatocellular carcinoma cells strain and also its effect on the proliferation, migration and adhesion of hepatocellular carcinoma cells. Methods: Three expression vectors of siRNA were constructed. Lipo2000 was employed to transfect Hca-F cells and Western blot was used to detect the inhibition effect of siRNA on the expression of Wisp-1 gene. Afterward, CCK8 was adopted to detect the effect of Wisp-1 siRNA on the proliferation of Hca-F cells; Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of Wisp-1 siRNA on the apoptosis of Hca-F cells; Transwell was used to detect the effect of Wisp-1 siRNA on the migration of Hca-F cells. The in vitro cell adhesion kit was used to detect of Wisp-1 siRNA on the change in the components of extracellular matrix to which Hca-F cells adhered. Western blot was used to detect the activation of protein kinase B (AKT)/glycogen synthase kinase-3β pathway and the expression of downstream target protein p53 and matrix metalloproteinases-2. Results: The siRNA showed interference effect on the expression of Wisp-1 gene. Compared with the control group, after being transfected to cells, Wisp-1 siRNA could significantly inhibit the proliferation, migration and adhesion of Hca-F cells and also promote the cell apoptosis, which was related to the down-regulated phosphorylation of AKT and glycogen synthase kinase-3β and the expression of p53 and matrix metalloproteinases-2 (P < 0.05). Conclusions: The inhibition of Wisp-1 expression can reduce the proliferation, migration and adhesion of mouse hepatocellular carcinoma cells, which is related to the AKT/glycogen synthase kinase-3β pathway. Wisp-1 gene may be the potential target to cure the hepatocellular carcinoma.
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<p><b>OBJECTIVE</b>To investigate the cell cycle changes of hepatoma cells and the effect of antisense oligonucleotide targeting bFGF on apoptosis in the hepatoma cells.</p><p><b>METHODS</b>The oligodeoxynucleotides were transfected with Lipofectin into hepatoma HepG2 cells. Inhibition of bFGF protein expression was assessed by confocal laser scanning microscopy and Western blot under the best condition of transfection of antisense oligonucleotide targeting bFGF, and the apoptosis in those cells was determined by flow cytometry. HepG2 cells were cultured in 24-well culture dish. The cultured cells were divided into 3 groups: group 1, the normal control group without any treatment; group 2, transfected with antisense oligonucleotide targeting bFGF; group 3, transfected with scrambled sequence targeting bFGF.</p><p><b>RESULTS</b>The results from confocal microscopy and Western blot showed an inhibition of expression of bFGF at different levels under the best condition of transfection with antisense oligonucleotide targeting bFGF. The treatment with antisense oligonucleotide of bFGF not only reduced the expression of bFGF revealed by confocal microscopy and Western blotting, but also increased the apoptosis in HepG 2 cells (P < 0. 01).</p><p><b>CONCLUSION</b>Treatment with antisense oligonucleotide of bFGF inhibits expression of bFGF protein and increase apoptosis. bFGF may take part in apoptosis regulation of hepatoma cells and may be used as a target in the treatment of hepatocellular carcinoma.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Cycle , Cell Line, Tumor , Fibroblast Growth Factor 2 , Genetics , Metabolism , Liver Neoplasms , Metabolism , Pathology , Oligonucleotides, Antisense , Pharmacology , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of Fuzheng Jiangnian Capsule (FZJN) on the pre-thrombosis correlated factors in patients with coronary heart disease (CHD).</p><p><b>METHODS</b>Ninety patients with CHD complicated with blood hyperviscosity syndrome were treated with conventional treatment and randomly divided into three groups by the additional treatment, i. e. the FZJN group (FZJN, a preparation with action of invigorating Pi, supplementing Shen, and activating blood circulation), the CSDP group [Compound Salviae droplet pill, CSDP) with the action of activating blood circulation to remove blood stasis) and the aspirin (ASP) group, 30 patients in each group. After two months of treatment, clinical efficacy, the levels of endothelin (ET), nitric oxide (NO), coagulation factor I (Fib), D-dimer (DD), thrombocytic granule membranous glucoprotein (CD62p), superoxide dismutase (SOD), high- and low-density lipoprotein-cholesterol (HDL-C, LDL-C) in patients before and after treatment were observed and compared with those in the healthy control group.</p><p><b>RESULTS</b>Compared with the healthy control group, the levels of Fib, DD, ET, CD62p were significantly higher, NO and SOD significantly lower (P<0.05, P<0.01) in all the patients with CHD. Compared with the same group before treatment, the levels of Fib, DD, ET, CD62p, LDL-C in the FZJN group lowered significantly, while NO and SOD raised significantly (P <0.05, P <0.01). ET and CD62p in the CSDP group lowered significantly, while SOD raised significantly (P < 0.05), CD62p in the ASP group lowered significantly (P < 0.05). No statistical difference was found in comparison of DD or ET, though certain improvement was shown. The total effective rate in relieving TCM syndromes and angina pectoris, and the decrease or stop rate of nitrate esters medication were superior in FZJN group to those in the CSDP group and the ASP group, respectively (P <0.05). The rate of electrocardiogram improvement in the FZJN and CSDP group was superior to that in the ASP group (P< 0.05).</p><p><b>CONCLUSION</b>Increase of Fib, DD, ET, CD62p, and decrease of NO and SOD levels were found in patients with CHD in prethrombosis stage. Compared with ASP, compound Chinese medicinal herbs can act on the prethrombosis manner of CHD patients through multi-paths, multi-links. FZJN showed better efficacy in improving correlated blood molecule markers and clinical syndromes than CSDP, suggesting that the possible mechanism of FZJN might be related to its actions in dilating blood vessels, improving microcirculation, alleviating endothelial cell damage, inhibiting activity of blood platelet, regulating coagulation-fibrolysis balance, improving metabolism of free radicals as well as lowering the level of LDL-C.</p>