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

Chatchai Phoomak, Dayoung Park, Atit Silsirivanit, Kanlayanee Sawanyawisuth, Kulthida Vaeteewoottacharn, Marutpong Detarya, Chaisiri Wongkham, Carlito B Lebrilla, Sopit Wongkham

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish (US)
JournalMolecular Oncology
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

Heterogeneous-Nuclear Ribonucleoprotein K
Cholangiocarcinoma
Neoplasm Metastasis
Proteins
Click Chemistry
Cell Line
Protein Databases
RNA-Binding Proteins
Survival
Cyclin D1
RNA Precursors
DNA-Binding Proteins
Post Translational Protein Processing
Immunoprecipitation
Proteomics
Small Interfering RNA

Keywords

  • bile duct cancer
  • heterogeneous nuclear ribonucleoprotein-K
  • metastasis
  • O-GlcNAcylated proteins

ASJC Scopus subject areas

  • Molecular Medicine
  • Genetics
  • Oncology
  • Cancer Research

Cite this

Phoomak, C., Park, D., Silsirivanit, A., Sawanyawisuth, K., Vaeteewoottacharn, K., Detarya, M., ... Wongkham, S. (Accepted/In press). O-GlcNAc-induced nuclear translocation of hnRNP-K is associated with progression and metastasis of cholangiocarcinoma. Molecular Oncology. https://doi.org/10.1002/1878-0261.12406

O-GlcNAc-induced nuclear translocation of hnRNP-K is associated with progression and metastasis of cholangiocarcinoma. / Phoomak, Chatchai; Park, Dayoung; Silsirivanit, Atit; Sawanyawisuth, Kanlayanee; Vaeteewoottacharn, Kulthida; Detarya, Marutpong; Wongkham, Chaisiri; Lebrilla, Carlito B; Wongkham, Sopit.

In: Molecular Oncology, 01.01.2019.

Research output: Contribution to journalArticle

Phoomak, C, Park, D, Silsirivanit, A, Sawanyawisuth, K, Vaeteewoottacharn, K, Detarya, M, Wongkham, C, Lebrilla, CB & Wongkham, S 2019, 'O-GlcNAc-induced nuclear translocation of hnRNP-K is associated with progression and metastasis of cholangiocarcinoma', Molecular Oncology. https://doi.org/10.1002/1878-0261.12406
Phoomak, Chatchai ; Park, Dayoung ; Silsirivanit, Atit ; Sawanyawisuth, Kanlayanee ; Vaeteewoottacharn, Kulthida ; Detarya, Marutpong ; Wongkham, Chaisiri ; Lebrilla, Carlito B ; Wongkham, Sopit. / O-GlcNAc-induced nuclear translocation of hnRNP-K is associated with progression and metastasis of cholangiocarcinoma. In: Molecular Oncology. 2019.
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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.",
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AU - Sawanyawisuth, Kanlayanee

AU - Vaeteewoottacharn, Kulthida

AU - Detarya, Marutpong

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