Establishing HEK-293T cell model carrying human sodium channel and PRKAG2 gene / 第二军医大学学报

ABSTRACT

Objective: To establish a HEK-293T cell model with sodium channel function similar to that in PRKAG2 syndrome. Methods: Vectors ofPRKAG2 gene and SCN5A gene were constructed and were used to co-transfect HEK-293T cells. Immunofluorescence, real-time PCR and Western blotting analysis were used to examine the transfection results and the expression of interested mRNA and protein. Results: Imunofluorescence findings showed red fluorescence and green fluorescence in HEK-293T cells 48 hours after co-transfection. Real-time PCR and Western blotting analysis showed SCN5A expression in pure SCN5A group, SCN5A + wild-type group, SCN5A + R302Q mutant group, and SCN5A + G100S mutant group, but not in the blank control group, showing significant significance (P<0.05). Expression of PRKAG2 gene and protein in SCN5A+PRKAG2 group was significantly higher than those in the blank control group, pure SCN5A group, SCN5A+R302Q, and SCN5A+G100S groups (P<0.05). Conclusion: We have successfully established a HEK-293T cell model with sodium channel function similar to that in PRKAG2 syndrome, paving a way for future study.