Objective:To establish a stable glycine receptor alpha 1(GlyRα1)-expressed HEK293 cell line. Methods:The pcDNA3.1-GlyRα1 plasmid was constructed and transfected into HEK293 cells with lipofectin. The stable transfectants were screened by G418. The mRNA expression of GlyRα1 was identified using RT-PCR. The protein expression was analyzed by Western blot. Results:Selected by G418, 4 out of 7 transfected cell lines showed high expression level of GlyRα1, as demonstrated by RT-PCR and Western blot analysis. No or low expression of GlyRα1 was shown in other 3 cell lines. Conclusion:A HEK293 cell line stably expressing GlyRα1 was constructed successfully.