Construction and identification of TRPM7 channel overexpression cell strain and preliminary functional research of the channel

To explore the function of TRPM7 (transient receptor potential melastatin 7) channel,the overexpression system of TRPM7 channel was constructed by transiently transfection of HEK293T (human embryonic kidney 293T) cell with TRPM7-eGFP plasmid.Through the fluorescence microscope analysis,the transfection efficiency was about 30%-40%.The corresponding expression of the gene and protein was observed by RT-PCR and western blotting. And the result of patch clamp showed that the whole cell current was about 2800pA.Acidic extracellular solution increased the TRPM7 inward currents and 200μmol/L 2-APB(2-aminoethyl diphenyl borate) inhibited the TRPM7 outward currents.These results demonstrated that the transiently transfected TRPM7-HEK293T cell line was successfully constructed.The acidic pH and the non-specific inhibitor of TRPM7,2-APB made different changes on TRPM7,which indicates that the function of the channel can be affected by multiple factors.