Relationship of Ctr1 mRNA Transcription Level to Intracellular Copper Ion Concentration

Objective To analyze the relationship of transcription level of copper transport protein 1(Ctr1)mRNA to the copper ion concentration in cells.Methods Ctr1 gene was amplified from subcultured porcine kidney PK15 cells,then cloned and sequenced.The effects of various copper ion concentrations(0,7.8,15.6,31.2 and 62.5 μmol /L)on the Ctr1 mRNA transcription level in PK15 cells were determined by semi-quantitative RT-PCR,using β-actin as an internal reference.The intracellular copper ion concentration was determined by atomic absorption spectroscopy.Results The homology of amplified Ctr1 gene to that reported in GenBank was 98%.The transcription level of Ctr1 mRNA showed a double-way response to the copper ion concentration at a range of 7.8 ～ 62.5 μmol /L in PK15 cells.The Ctr1 mRNA transcription levels of various test groups were lower than that in control group(copper ion concentration 0 μmol /L).However,the intracellular copper ion concentration in test groups were higher than that in control group,of which that in group Ⅳ(copper ion concentration 31.2 μmol /L)was significantly higher than those in the other groups.Conclusion The transcription level of Ctr1 mRNA was negatively related to intracellular copper ion concentration.