title: Quantitative glycome analysis of N-glycan patterns in bladder cancer vs normal bladder cells using an integrated strategy.
abstract: Diagnosis of bladder cancer, one of the most common types of human cancer, at an early (nonmuscle-invasive) stage is the best way to reduce the mortality rate. Tumor malignancy in general is closely associated with alterations of glycan expression. Glycosylation status, particularly global glycomes, in bladder cancer has not been well studied. We integrated lectin microarray and mass spectrometry (MS) methods to quantitatively analyze and compare glycan expression in four bladder cancer cell lines (KK47, YTS1, J82, T24) and one normal bladder mucosa cell line (HCV29). Glycopattern alterations were analyzed using lectin microarray analysis and confirmed by lectin staining and lectin blotting. Associations of glycopatterns with diverging stages were evaluated by lectin histochemistry on tissue microarrays. N-Glycans were derivatized by amidation of sialylated glycans with acetohydrazide and reductive amination with the stable isotope tags [(12)C6]- and [(13)C6]-aniline, and were quantitatively analyzed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). N-Glycan biosynthesis-associated proteins were quantitatively analyzed by a stable isotope labeling by amino acids in cell culture (SILAC) proteomics method, which revealed significant differences in expression of 13 glycosyltransferases and 4 glycosidases. Our findings indicate that sialyl Lewis X (sLe(x)), terminal GalNAc and Gal, and high mannose-type N-glycans were more highly expressed in bladder cancer cells and tissues than in normal cells. Bladder cancer cells showed high expression of core-fucosylated N-glycans but low expression of terminally fucosylated N-glycans. Each of these glycome changes may be directly related to bladder cancer progression.
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education; School of Biotechnology, Jiangnan University , Wuxi, China.
authors.forename: Ganglong
authors.initials: G
authors.lastname: Yang
authors.forename: Zengqi
authors.initials: Z
authors.lastname: Tan
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.forename: Jia
authors.initials: J
authors.lastname: Guo
authors.forename: Hanjie
authors.initials: H
authors.lastname: Yu
authors.forename: Jingmin
authors.initials: J
authors.lastname: Yu
authors.forename: Chengwen
authors.initials: C
authors.lastname: Sun
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Qi
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Guan
mesh_terms: D002241:Carbohydrates / Q000737:chemistry*; D002460:Cell Line; D045744:Cell Line, Tumor; D006801:Humans; D011134:Polysaccharides / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D001743:Urinary Bladder / Q000166:cytology / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology
chemical_list: D002241:Carbohydrates; D011134:Polysaccharides
keywords: N-glycans; SILAC method; bladder cancer; lectin microarray analysis; mass spectrometry; quantitative glycomics