title: Proteomic analysis and abrogated expression of O-GlcNAcylated proteins associated with primary breast cancer.
abstract: O-GlcNAcylation is a dynamic PTM of nuclear and cytoplasmic proteins, regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase, which catalyze the addition and removal of O-GlcNAc, respectively. This modification is associated with glucose metabolism, which plays important roles in many diseases including cancer. Although emerging evidence reveals that some tumor-associated proteins are O-GlcNAc modified, the total O-GlcNAcylation in cancer is still largely unexplored. Here, we demonstrate that O-GlcNAcylation was increased in primary breast malignant tumors, not in benign tumors and that this augmentation was associated with increased expression of OGT level. Using 2D O-GlcNAc immnoblotting and LC-MS/MS analysis, we successfully identified 29 proteins, with seven being uniquely O-GlcNAcylated or associated with O-GlcNAcylation in cancer. Of these identified proteins, some were related to the Warburg effect, including metabolic enzymes, proteins involved in stress responses and biosynthesis. In addition, proteins associated with RNA metabolism, gene expression, and cytoskeleton were highly O-GlcNAcylated or associated with O-GlcNAcylation. Moreover, OGT knockdown showed that decreasing O-GlcNAcylation was related to inhibition of the anchorage-independent growth in vitro. These data indicate that aberrant protein O-GlcNAcylation is associated with breast cancer. Abnormal modification of these O-GlcNAc-modified proteins might be one of the vital malignant characteristics of cancer.
authors.affiliation: Laboratory of Biochemistry, Chulabhorn Research Institute, Bangkok, Thailand. voraratt@cri.or.th
authors.forename: Voraratt
authors.initials: V
authors.lastname: Champattanachai
authors.forename: Pukkavadee
authors.initials: P
authors.lastname: Netsirisawan
authors.forename: Parunya
authors.initials: P
authors.lastname: Chaiyawat
authors.forename: Thanong
authors.initials: T
authors.lastname: Phueaouan
authors.forename: Ratana
authors.initials: R
authors.lastname: Charoenwattanasatien
authors.forename: Daranee
authors.initials: D
authors.lastname: Chokchaichamnankit
authors.forename: Phaibul
authors.initials: P
authors.lastname: Punyarit
authors.forename: Chantragan
authors.initials: C
authors.lastname: Srisomsap
authors.forename: Jisnuson
authors.initials: J
authors.lastname: Svasti
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry; D014408:Biomarkers, Tumor; D001943:Breast Neoplasms / Q000737:chemistry* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D015151:Immunoblotting; D019344:Lactic Acid; D008954:Models, Biological; D017351:N-Acetylglucosaminyltransferases; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D014408:Biomarkers, Tumor; D006023:Glycoproteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D020543:Proteome; D019344:Lactic Acid; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase; D000117:Acetylglucosamine
keywords: Breast cancer; Glycoproteomics; Hexosamine biosynthesis pathway; O-GlcNAc transferase; O-GlcNAcyla-tion; Warburg effect