【摘要】 [目的]研究镉对人胚肾细胞(HEK293T)金属硫蛋白(MT)各基因亚型表达的影响。[方法]以10μmol/LCdCl2染毒HEK 293T0、2、4、8、12、16、20、24h,2.5、5、10、20、40μmol/LCdCl2染毒HEK293T24h后,采用MTT法测定HEK 293T增殖活性;采用RT-PCR检测镉对HEK 293T MT-1A、MT-1B、MT-1E、MT-1F、MT-1G、MT-1H、MT-1X及MT-2A mRNA表达的影响。[结果]CdCl2染毒HEK 293T 24h后,经10μmol/LCdCl2诱导后,随时间MT-1A、MT-1F、MT-1G、MT-1H及MT-2A mRNA表达水平均呈先升高后降低趋势。MT-1X mRNA水平在8h明显升高(P<0.05),16h达到峰值并持续到24h。5μmol/L以上CdCl2均可明显抑制HEK293T活力,与对照组相比差异均有统计学意义(P<0.05)。MT-1A、MT-1F、MT-1G、MT-1H及MT-2A mRNA在2.5μmol/LCdCl2诱导表达水平均达到峰值,随着剂量增加,MT-1F、MT-1G、MT-1H维持在较高水平,40μmol/L剂量水平仍明显高于对照组(P<0.05),MT-1A、MT-2A在40μmol/L降低到基础表达水平。MT-1X mRNA表达水平与染毒剂量呈线性正相关(r=0.75,P<0.01)。MT-1X的表达与镉对HEK 293T毒性呈负相关关系(r=-0.88,P<0.01)。[结论]镉可以诱导MT-1F、MT-1G、MT-1H、MT-1X及MT-2A mRNA表达水平明显升高,并表现出特定的剂量-效应和时间-效应关系。更多还原

【Abstract】 [ Objective] To explore gene expression of metallothionein 1(MT-1)isoforms in human embryo kidney cel(lHEK 293T).[ Methods] Treatment with 2.5, 5, 10, 20, and 40 μmol/L CdCl2 for 24 h, and 10 μmol/L CdCl2 for 0, 2, 4, 8, 12, 16, 20, and 24 h to HEK 293T cell, the cell viability was measured with MTT assay. The expression of MT-1A, MT-1B, MT-1E, MT-1F, MT-1G, MT-1H, MT-1X and MT-2A mRNA was determined in HEK 239T cell after exposure to cadmium by quantitative RT-PCR. [Results] Treatment with different levels of CdCl2 for 24 h significantly reduced the viability in more than 5 μmol/L CdCl2. The diverse expression patterns of these genes(MT-1A, MT-1F, MT-1G, MT-1H, MT-1X and MT-2A) mRNA expression depending on Cd concentration and exposure time was significantly increased after exposure to cadmium(P < 0.05).[ Conclusion] Results indicated that MT-1A, MT-1E, MT-1F, MT-1X, and MT-2A mRNA expression was induced by cadmium, and exhibits dose-effect and dose-course relationship with Cd exposure level.更多还原