ABSTRACT
Objective To study the influence of long time exposure to low levels of sodium arsenite on Warburg effect in cultured human epithelial cells (SV-HUC-1) at different times.Methods SV-HUC-1 cells were exposed to 0.5 μmol/L sodium arsenite for 10,20,30 weeks and cells cultured without sodium arsenite for 10,20,30 weeks were regarded as control groupsin vitro.Lactate assay kit and glucose assay kit were used to measure the lactate secretion and glucose consumption levels,and cells mRNA and protein expressions of SCL2A1 and hexokinase2(HK2) were detected using Real-time PCR and Western blotting.Results The levels of lactate secretion [(4.67 ± 0.20),(7.47 ± 0.28),(12.46 + 0.47) mmol/L],glucose consumption [(2.86 ± 0.11),(4.25 ± 0.19),(6.38 ± 0.05) mmol/L] and expression of HK2 protein (1.21 ± 0.06,1.36 ± 0.13,1.60 ± 0.12) increased significantly after treated with 0.5 μmol/L sodium for 10,20,30 weeks compared with those of control groups [(3.04 ± 0.11),(3.90 ± 0.32),(4.77 ± 0.24) mmol/L;(2.17 ± 0.15),(2.48 ± 0.24),(2.71 ± 0.13) mmol/L;1.00 ± 0.00;P ＜ 0.05].Compared with control group,the expressions of SCL2A1 mRNA,HK2 mRNA and SCL2A1 protein in SV-HUC-1 cells treated with sodium arsenite for 10 weeks increased but the difference was not statistically significant (P ＞ 0.05).While the expressions of SCL2A1 mRNA,HK2 mRNA and SCL2A1 protein in SV-HUC-1 cells treated with sodium arsenite for 20 and 30 weeks increased significantly compared to those of control groups (P ＜ 0.05).Conclusion Long-term exposure to low concentrations of sodium arsenite can increase glycolysis in SV-HUC-1 and induce Warburg effect.
ABSTRACT
Objective To study the state of oxidative stress induced by sodium arsenite (NaAsO2) in SV-HUC-1 cell. Methods MTT assay was used to evaluate the viability of the cells. The level of ROS was detected by staining cells with DCFH-DA. The content of GSH and MDA were measured by DTNB and thiobarbituric acid methods. The activity of SOD was measured by xanthine oxidase method. Results Compared with the control group, the viability cells decreased in all the treated groups (P0.05), the activity of SOD in all the treated groups was significantly decreased (P
ABSTRACT
Although protein kinase B (PKB, AKT) has been investigated extensively for its roles in oncogenic transformation and apoptotic prevention, controversial results are also reported. Here we assessed the role of AKT in the cell growth and expression of a key set of cell cycle regulators in the human normal uroepithelial cell line, SVHUC-1. AKT activity was suppressed by permanent transfection of dominent negative (DN)-AKT with Lipofectamine Plus. Cell viability was measured by the crystal violet assay. DNA contents stained by propidium iodide were measured by flow-cytometry for cell cycle analysis. Cell growth curve showed that overexpression of DN-AKT which suppressed the AKT activity decreased the cell growth. In the cell cycle analysis, overexpression of DN-AKT resulted in a 6% increase in the proportion of cells in G1 phase. Furthermore, DN-AKT overexpression increased the p27(Kip1) protein expression and the activation of a transcription factor FKHR, which induces p27(Kip1) transcription. Our results suggest that, in normal uroepithelial cells, AKT activation increases the cell growth through the influence on p27(Kip1) expression and FKHR activation. Thus, AKT may be used as a biomarker for tumor transformation of bladder uroepithelial cells.