Human gastric cancer is a big public health problem.Multidrug resistance is a main obstacle to successful chemotherapeutic treatment in gastric cancers and the underlying mechanism is not clear.Glycosylation,one of the most important post translational modifications of proteins,plays a vital role in diverse aspects of tumor progression.In the present study,we applied two multidrug resistance cell lines and their parental drug sensitive gastric cancer cell line to a modified cell surface capturing strategy with triplex labeling to characterize MDR related cell surface glycoproteome.Finally,56 cell membrane glycoproteins were successfully identified via combination of identification by glycopeptides and quantitation by non-glycopeptides,and 11 of them were found to be differentially expressed with the same trend in both drug resistant cell lines compared with that in sensitive cell line.The further analysis by western blot and in vitro drug sensitivity assay demonstrated that our approach is reliable and accurate and suggested that these glycoproteins may represent as biomarkers for multidrug resistance in gastric cancer.