Objective To confirm the enhancing effect of excision repair cross complementing rodent repair deficiency gene 2 (ERCC2) on alkylating agents resistance. Methods The authors constructed a pcDNA3-ERCC2 plasmid. The pcDNA3-ERCC2 was transfected into a selected ERCC2 negative human glioma cell line,SKMG-4,using liposome-mediated transfection. After G418 selection,a stable transfected cell line was obtained and tested for cytotoxicity of several alkylating agents.Results The stable transfectant was obtained and confirmed by RT-PCR as well as Western blot analysis to be strongly expressing ERCC2 at both mRNA and protein levels. The IC 90 (μmol/L) of two alkylating agents,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 that ERCC2 contributes,at least partially,to alkylating agent resistance in human glioma cell line. Methods The authors constructed a pcDNA3-ERCC2 plasmid. The pcDNA3-ERCC2 was transfected into a selected ERCC2 negative human glioma cell line, SKMG-4, using liposome-mediated transfection. After G418 selection, a stable transfected cell line was obtained and tested for cytotoxicity of several alkylating agents. Results of the stable transfectant was obtained and confirmed by RT-PCR as well as Western blot analysis to be strongly expressing ERCC2 at both mRNA and protein levels. respectively The IC 90 (μmol / L) of two alkylating agents, cisplatin and melphalan, increased from 1.0 to 1.75 (75%) and 5.6 to 9.0 cell line. Conclusion The present data provided evidences and confirmed the authors’ previous results that ERCC2 contributes, at least partially, to alkylating agent resistance in human glioma cell line.