Title: Expression of MDR1,GST-π and Arsenic Tolerance of L-02 Cell after 6 Weeks Culture with Low Concentration of Arsenic
Abstract: [Objective]To study the expression of multidrug resistance 1(MDR1)and glutathione S-transferase-π(GST-π) in acquired arsenic tolerant L-02 hepatic cells,and to discuss its relationship with the acquired tolerance to arsenic.[Methods] MTT(3-[4,5-dimethy thiazol-2-yl]-2,5-diphenyl tetrazolium)assay was conducted to detect the effects of NaAsO2 exposure in various concentrations(0,1,10,20,30,40,50,60,70,80,90,and 100 μmol/L)for 24 h on the survival rate of L-02 cells,then the initial dose under which the cell survival rate was 90%-95% was chosen to be added into the medium for culturing experiment L-02 cells(induced cells).L-02 cells grown in medium without arsenic were provided as contro(lnormal cells).Induced cells and normal cells were cultured under the same condition for 6 weeks,then the survival rate and LC50 of induced cells and normal cells were detected by MTT assay which can reflect the change of arsenic tolerance.The levels of MDR1 and GST-π mRNA were determined by real-time quantitative PCR,and the expression of P-glycoprotein(P-gp),GST-π was examined by immunohistochemical SABC.Cell survival rate and intracellular concentrations of total arsenic of the cells that were cultured with 10 μmol/L NaAsO2 for 24 h were measured in order to observe the response of the cells to another large dose acute arsenic exposure.[Results] After 6 weeks after induction of NaAsO2,LC50 and survival rate of induced cells were significantly higher than that of normal cells(P0.001);expression of the genes for GST-π/MDR1 in induced cells was significantly higher than that in normal cells(P0.001).Compared with normal cells,expression of P-pg/GST-π protein in induced cells was significantly higher(P0.001).In the re-trial of acute arsenic poisoning,the survival rate of induced cells was significantly higher than that of normal cells(P0.001),the total cellular arsenic content was markedly decreased in induced cells(P0.001).[Conclusion] L-02 cells possess induced arsenic tolerance phenomenon.Acquired tolerance to arsenic is associated with increased expression of MDR1/GST-π in human hepatocyte cell line.This may provide an experimental basis for the prevention and treatment of arsenic poisoning at the gene level.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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