O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma
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 p...
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doaj-482137518b9d4e0287a97bc77c3193402020-11-25T03:42:28ZengWileyMolecular Oncology1574-78911878-02612019-02-0113233835710.1002/1878-0261.12406O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinomaChatchai Phoomak0Dayoung Park1Atit Silsirivanit2Kanlayanee Sawanyawisuth3Kulthida Vaeteewoottacharn4Marutpong Detarya5Chaisiri Wongkham6Carlito B. Lebrilla7Sopit Wongkham8Department of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Surgery Beth Israel Deaconess Medical Center Harvard Medical School Boston MA USADepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandDepartment of Chemistry University of California Davis CA USADepartment of Biochemistry Faculty of Medicine Khon Kaen University ThailandO‐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.https://doi.org/10.1002/1878-0261.12406bile duct cancerheterogeneous nuclear ribonucleoprotein‐KmetastasisO‐GlcNAcylated proteins |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chatchai Phoomak Dayoung Park Atit Silsirivanit Kanlayanee Sawanyawisuth Kulthida Vaeteewoottacharn Marutpong Detarya Chaisiri Wongkham Carlito B. Lebrilla Sopit Wongkham |
spellingShingle |
Chatchai Phoomak Dayoung Park Atit Silsirivanit Kanlayanee Sawanyawisuth Kulthida Vaeteewoottacharn Marutpong Detarya Chaisiri Wongkham Carlito B. Lebrilla Sopit Wongkham O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma Molecular Oncology bile duct cancer heterogeneous nuclear ribonucleoprotein‐K metastasis O‐GlcNAcylated proteins |
author_facet |
Chatchai Phoomak Dayoung Park Atit Silsirivanit Kanlayanee Sawanyawisuth Kulthida Vaeteewoottacharn Marutpong Detarya Chaisiri Wongkham Carlito B. Lebrilla Sopit Wongkham |
author_sort |
Chatchai Phoomak |
title |
O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma |
title_short |
O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma |
title_full |
O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma |
title_fullStr |
O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma |
title_full_unstemmed |
O‐GlcNAc‐induced nuclear translocation of hnRNP‐K is associated with progression and metastasis of cholangiocarcinoma |
title_sort |
o‐glcnac‐induced nuclear translocation of hnrnp‐k is associated with progression and metastasis of cholangiocarcinoma |
publisher |
Wiley |
series |
Molecular Oncology |
issn |
1574-7891 1878-0261 |
publishDate |
2019-02-01 |
description |
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. |
topic |
bile duct cancer heterogeneous nuclear ribonucleoprotein‐K metastasis O‐GlcNAcylated proteins |
url |
https://doi.org/10.1002/1878-0261.12406 |
work_keys_str_mv |
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