ABSTRACT
To explore the effect and mechanism of quercetin on proliferation and apoptosis of leukemia cells, and provide a theoretical basis for its clinical application. HL-60 leukemia cell lines was treated with different dose quercetin, the proliferation activity of leukemia cells was assessed by MTT method; the morphological changes of apoptosis of HL-60 cells, including nuclear condensation and DNA fragmentation, were observed by Hoechst 33258 fluorescence staining, the apoptosis rate and caspase 2,3 activation were assessed by flow cytometry, and the cell signal pathway including phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (pAkt), Bcl-2, Bax were detected by western blotting. Quercetin could significantly decrease the proliferation activity of HL-60 cells through the blockade of G(0)/G(1) phase, and induce the apoptosis of HL-60 cells in a time- and dose-dependent manner. Quercetin caused leukemia cells apoptosis by decreasing the protein expression of PI3K and Bax, the inhibitory phosphorylation of Akt, the decreased levels of Bcl-2 protein and increased activations of caspase-2 and -3, and increased poly(ADP-ribose) polymerase cleavage. Our results indicate that the apoptotic processes caused by quercetin are mediated by the decrease of pAkt and Bcl-2 levels, the increase of Bax level, and the activation of caspase families in HL-60 cells.
Subject(s)
Apoptosis/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Quercetin/pharmacology , bcl-2-Associated X Protein/biosynthesis , Caspase 2/biosynthesis , Caspase 2/drug effects , Caspase 3/biosynthesis , Caspase 3/drug effects , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , DNA Fragmentation/drug effects , HL-60 Cells , Humans , Leukemia/metabolism , Phosphorylation/drug effects , Poly(ADP-ribose) Polymerases/metabolism , Signal Transduction/drug effectsABSTRACT
Objective To investigate the expression of Oct4 in liver cancer, and the interrelation of the Oct4 and Wnt/β-catenin genes in hepatocellular carcinoma( HCC) cell line HepG2. Methods RTPCR technique was used to detect the expression of Oct4 and β-Catenin in HCC specimens; RNAi was used to knock-down the expression of Oct4 in HepG2, and the change of Wnt/β-catenin related genes were detected by Real time-PCR. Results In HCC specimens, the expression of Oct4 and β-Catenin in tumor and cirrhotic liver tissues were stronger than normal liver tissues. In SiRNA Oct4 HepG2 cells, the expression of Oct4 was downregulated, and β-catenin as well as Wnt10b were in a positive correlation with Oct4, TCF3 was in negative correlation with Oct4. Clone formation and move ability of the HepG2 were downregulated. Conclusions The expression of Oct4 was higher in tumor tissues than in normal liver tissues. Silencing Oct4 by SiRNA-0ct4 in HepG2 resulted in decreased ability of clone formation and cell movement.
ABSTRACT
Objective To study the relationship between serum leptin and fibrosis in chronic hepatitis B incorporate alcoholic liver disease(CHB + ALD) patients.Methods Select 17 male patients with CHB+ALD,15 male patients with liver cirrhosis(LC) ,19 male patients with chronic hepatitis B(CHB) and 12 male healthy persons as normal control(NC).Serum leptin is detected by ELISA in 63 patients.Serum levels of fibrosis markers(HA,LN,PC HI ,IV-C) are determined at the same time.Results Serum leptin level in CHB +ALD are significantly higher than those of the NC(6.79±24.12) μg/L vs (4.27±7.18 ) μg/L (P＜0.05).There is no significant difference between patients with CHB and the NC.The four fibrosis markers in CHB + ALD patients are significantly higher than that of men both in the CHB and the NC.There are significant correlations between both serum leptin level and quadrinomial fibrosis markers(P＜0.05 or P＜0.01 ).Conclusion That serum leptin level is one aecelerate gene in the fibrosis formation in patients with CHB+ALD.
ABSTRACT
Objective To prepare cytokine-induced killer(CIK) in vitro, and study the biological activities of CIK. Methods Lymphocytes were isolated from peripheral blood and umbilical blood, and induced by cytokine.The cell surface markers CD_3 and CD_~56 of CIK were analyzed by FACS. The proliferation of CIK was tested using 3H -TdR release method, and the cytotoxic effect of CIK on hepatic carcinoma cells was measured by MTT. Results CIK cells quickly proliferated from the second week, and expanded more than 60-fold at the 31th day after induction. The CD_3+ and CD_~56 + cells expanded more than 800 folds. The cytotoxic effect of CIK on hepatic carcinoma cells was obviously stronger than that of LAK cells, and CIK had not obvious cytotoxic effect on fetal liver cells. Conclusion CIK was a highly efficient cytotoxic cells against tumors, and had clinical application potentials.
