O-GlcNAc-induced nuclear translocation of hnRNP-K is associated with progression and metastasis of cholangiocarcinoma.

Abstract

O-GlcNAcylation is a key post-translational modification that modifies the functions of proteins. Associations between O-GlcNAcylation, shorter survival of cholangiocarcinoma (CCA) patients, and increased migration/invasion of CCA cell lines have been reported. However, the specific O-GlcNAcylated proteins (OGPs) that participate in promotion of CCA progression are poorly understood. OGPs were isolated from human CCA cell lines, KKU-213 and KKU-214, using a click chemistry-based enzymatic labeling system, identified using LC-MS/MS, and searched against an OGP database. From the proteomic analysis, a total of 21 OGPs related to cancer progression were identified, of which 12 have not been previously reported. Among these, hnRNP-K, a multifaceted RNA- and DNA-binding protein known as a pre-mRNA-binding protein, was one of the most abundantly expressed, suggesting its involvement in CCA progression. O-GlcNAcylation of hnRNP-K was further verified by anti-OGP/anti-hnRNP-K immunoprecipitations and sWGA pull-down assays. The perpetuation of CCA by hnRNP-K was evaluated using siRNA, which revealed modulation of cyclin D1, XIAP, EMT markers, and MMP2 and MMP7 expression. In native CCA cells, hnRNP-K was primarily localized in the nucleus; however, when O-GlcNAcylation was suppressed, hnRNP-K was retained in the cytoplasm. These data signify an association between nuclear accumulation of hnRNP-K and the migratory capabilities of CCA cells. In human CCA tissues, expression of nuclear hnRNP-K was positively correlated with high O-GlcNAcylation levels, metastatic stage, and shorter survival of CCA patients. This study demonstrates the significance of O-GlcNAcylation on the nuclear translocation of hnRNP-K and its impact on the progression of CCA.

• Authors: Chatchai Phoomak, Dayoung Park, Atit Silsirivanit, Kanlayanee Sawanyawisuth, Kulthida Vaeteewoottacharn, Marutpong Detarya, Chaisiri Wongkham, Carlito B Lebrilla, Sopit Wongkham
• Journal: Molecular oncology (Mol Oncol) Vol. 13 Issue 2 Pg. 338-357 (02 2019) ISSN: 1878-0261 [Electronic] United States
• PMID: 30444036 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.

Chemical References

• Biomarkers, Tumor
• Heterogeneous-Nuclear Ribonucleoprotein K
• Glucosamine

Topics

• Bile Duct Neoplasms (metabolism, pathology)
• Biomarkers, Tumor (metabolism)
• Cell Line, Tumor
• Cell Movement
• Cell Nucleus (metabolism)
• Cell Proliferation
• Cholangiocarcinoma (metabolism, pathology)
• Disease Progression
• Female
• Glucosamine (metabolism)
• Glycosylation
• Heterogeneous-Nuclear Ribonucleoprotein K (metabolism)
• Humans
• Male
• Middle Aged
• Multivariate Analysis
• Neoplasm Invasiveness
• Neoplasm Metastasis
• Protein Transport
• Treatment Outcome