镉诱导人胚肾细胞金属硫蛋白基因亚型的表达
Metallothion Isoforms Gene Expression Induced by Cadmium in Human Embryo Kidney Cell
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摘要:
目的 研究镉对人胚肾细胞(HEK 293T)金属硫蛋白(MT)各基因亚型表达的影响。 方法 以10 μmol/LCdCl2染毒HEK 293T 0、2、4、8、12、16、20、24 h,2.5、5、10、20、40 μmol/L CdCl2染毒HEK 293T 24 h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 24 h后,经10 μmol/L CdCl2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8 h明显升高(P<0.05),16 h达到峰值并持续到24 h。5 μmol/L以上CdCl2均可明显抑制HEK 293T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2.5 μmol/L CdCl2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,40 μmol/L剂量水平仍明显高于对照组(P<0.05),MT-1A、MT-2A在40 μmol/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。 Abstract:Objective To explore gene expression of metallothionein 1 (MT-1)isoforms in human embryo kidney cell(HEK 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.
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