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申旭波, 周远忠, 姜慧, 贾飞飞, 熊云刚, 邹焰. 多药耐药基因1与人肝细胞耐砷性关系[J]. 中国公共卫生, 2009, 25(10): 1212-1214. DOI: 10.11847/zgggws2009-25-10-32
引用本文: 申旭波, 周远忠, 姜慧, 贾飞飞, 熊云刚, 邹焰. 多药耐药基因1与人肝细胞耐砷性关系[J]. 中国公共卫生, 2009, 25(10): 1212-1214. DOI: 10.11847/zgggws2009-25-10-32
SHEN Xu-bo, ZHOU Yuan-zhong, JIANG Hui, . Overexpression of MDR1 is associated with acquired tolerance to arsenic in L-02 cells[J]. Chinese Journal of Public Health, 2009, 25(10): 1212-1214. DOI: 10.11847/zgggws2009-25-10-32
Citation: SHEN Xu-bo, ZHOU Yuan-zhong, JIANG Hui, . Overexpression of MDR1 is associated with acquired tolerance to arsenic in L-02 cells[J]. Chinese Journal of Public Health, 2009, 25(10): 1212-1214. DOI: 10.11847/zgggws2009-25-10-32

多药耐药基因1与人肝细胞耐砷性关系

Overexpression of MDR1 is associated with acquired tolerance to arsenic in L-02 cells

  • 摘要: 目的观察多药耐药基因1(MDR1)在耐砷的人肝细胞(L-02)中的表达情况,探讨其与L-02耐砷性的关系。方法培养L-02和耐砷的L-02,用real-time PCR和免疫组织化学法检测细胞MDR1 mRNA和P糖蛋白的表达情况;2种细胞单用亚砷酸钠(NaAsO2)2.5,5.0,10 mmol/L及NaAsO2与MDR1抑制剂环胞菌素D衍生物(PSC833)联合作用24 h后收集细胞,四甲基偶氮噻唑蓝(MTT)法和石墨炉原子吸收光谱法检测细胞生存率和细胞内总砷浓度。结果MDR1 mRNA及P糖蛋白在耐砷的L-02中的表达强度分别为(8.890±0.968),(68.76±3.81)%,高于L-02的(1.014±0.189),(24.44±4.03)%,差异有统计学意义(P<0.001);NaAsO2单用时,低、中、高剂量组耐砷的L-02生存率分别为(109.630±0.511)%,(95.469±0.054)%,(78.890±0.024)%,明显高于L-02的(104.841±0.015)%,(91.534±0.026)%,(64.811±0.079)%;而耐砷的L-02内总砷浓度分别为(0.682±0.004),(1.355±0.005),(3.274±0.010)μg/L,明显低于L-02的(1.165±0.010),(3.661±0.002),(7.529±0.006)μg/L,差异有统计学意义(P<0.001);NaAsO2与PSC833联合作用则可增加细胞内总砷浓度、降低细胞存活率(P<0.001)。结论L-02细胞的耐砷性与MDR1高表达有关,在mRNA和蛋白水平均有体现。

     

    Abstract: ObjectiveTo study the relativity of multidrug resistance gene 1(MDR1)gene expression and acquired to lerance to arsenic in human hepatocyte cell line(L-02).MethodsThe levels of MDR1 mRNA were determined by real time quantitative PCR,and the expression of P-gly coprote in(P-gp)was examined by mimunohisto chemical SABC in both arsenite exposed cells and the control.Then the cells were incubated with fresh medium containing 0,2 5,5 0,10.0 mol/L NaAsO2 in the pre sence or absence of 10 mol/L PSC833(an inhibitor for Pgp).Twenty four hours later,cellular total arsenic was determined using graphite furnace atomic absorption spectrometry,and 3-4,5-dimethy lthiazol-2-yl-2,5-diphenyl tetrzolium(MTT)was used to detect cell survival rate.ResultsThe expression of MDR 1 mRNA and P-gp in arsenite exposed cells(8.890±0.968,68.76±3.81%)was higher than in L-02 cells(1.014±0.189,24.44±4.03%), (P<0.001);the survival rate of arsenite exposed L-02 cells(109.630±0.511%,95.469±0.054%,78.890±0.024%) was apparently higher than that of L-02 cells(104.841±0.015%,91.534±0.026%,64.811±0.079%),(P<0.001); cellular arsenic content in arsenite exposed cells(0.68±3.65×10-03,1.36±4.72×10-03,3.27±9.62×10-03 μg/L)was markedly decreased than in control(1.17±5.48×10-04,3.66±1.95×10-03,7.53±6.20×10-03 μg/L).With 10 mol/L PSC 833,cellular arsenic content was markedly increased and survival rate of the cells was apparently decreased (P<0.001).ConclusionAcquired to lerance to arsenic is associated with increased expression of MDR1 with presentation of mRNA and prote in in human hepatocyte cell line.

     

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