Objective To confirm the enhancing effect of excision repair cross complementing rodent repairdeficiency gene 2 (ERCC2) on alkylating agents resistance. Methods The authors constructed a pcDNA3-ERCC2 plasmid. The pcDNA3-ERCC2 was transfected into aselected ERCC2 negative human glioma cell line, SKMG-4, using liposome-mediated transfection. AfterG418 selection, a stable transfected cell line was obtained and tested for cytotoxicity of several alkylatingagents.Results The stable transfectant was obtained and confirmed by RT-PCR as well as Western blot analysisto be strongly expressing ERCC2 at both mRNA and protein levels. The 1O90 (pmol/L) of two alkylatingagents, cisplatin and melphalan, increased from 1.0 to 1.75 (75%) and 5.6 to 9.0 (61%), respectively,compared with control cell line.Conclusion The present data provided evidences and confirmed the authors'previous results thatERCC2 contributes, at least partially, to alkylating agent resistance in human glioma cell line.