Cargando…

Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line

Neuroblastoma (NB) development and progression are accompanied by changes in N-glycans attached to proteins. Here, we investigated the role of N-acetylglucosaminyltransferase-II (GnTII, MGAT2) protein substrates in neuroblastoma (NB) cells. MGAT2 was silenced in human BE(2)-C NB (HuNB) cells to gene...

Descripción completa

Detalles Bibliográficos
Autores principales: Hall, M. Kristen, Whitman, Austin A., Weidner, Douglas A., Schwalbe, Ruth A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7236022/
https://www.ncbi.nlm.nih.gov/pubmed/32260356
http://dx.doi.org/10.3390/biology9040071
_version_ 1783536084769046528
author Hall, M. Kristen
Whitman, Austin A.
Weidner, Douglas A.
Schwalbe, Ruth A.
author_facet Hall, M. Kristen
Whitman, Austin A.
Weidner, Douglas A.
Schwalbe, Ruth A.
author_sort Hall, M. Kristen
collection PubMed
description Neuroblastoma (NB) development and progression are accompanied by changes in N-glycans attached to proteins. Here, we investigated the role of N-acetylglucosaminyltransferase-II (GnTII, MGAT2) protein substrates in neuroblastoma (NB) cells. MGAT2 was silenced in human BE(2)-C NB (HuNB) cells to generate a novel cell line, HuNB(-MGAT2), lacking complex type N-glycans, as in rat B35 NB cells. Changes in N-glycan types were confirmed by lectin binding assays in both cell lines, and the rescued cell line, HuNB(-/+MGAT2). Western blotting of cells heterologously expressing a voltage-gated K+ channel (Kv3.1b) showed that some hybrid N-glycans of Kv3.1b could be processed to complex type in HuNB(-/+MGAT2) cells. In comparing HuNB and HuNB(-MGAT2) cells, decreased complex N-glycans reduced anchorage-independent cell growth, cell proliferation, and cell invasiveness, while they enhanced cell-cell interactions. Cell proliferation, invasiveness and adhesion of the HuNB(-/+MGAT2) cells were more like the HuNB than HuNB(-MGAT2). Western blotting revealed lower protein levels of MMP-2, EGFR and Gab2 in glycosylation mutant cells relative to parental cells. Gelatin zymography demonstrated that decreased MMP-2 protein activity was related to lowered MMP-2 protein levels. Thus, our results support that decreased complex type N-glycans suppress cell proliferation and cell invasiveness in both NB cell lines via remodeling ECM.
format Online
Article
Text
id pubmed-7236022
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-72360222020-05-28 Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line Hall, M. Kristen Whitman, Austin A. Weidner, Douglas A. Schwalbe, Ruth A. Biology (Basel) Article Neuroblastoma (NB) development and progression are accompanied by changes in N-glycans attached to proteins. Here, we investigated the role of N-acetylglucosaminyltransferase-II (GnTII, MGAT2) protein substrates in neuroblastoma (NB) cells. MGAT2 was silenced in human BE(2)-C NB (HuNB) cells to generate a novel cell line, HuNB(-MGAT2), lacking complex type N-glycans, as in rat B35 NB cells. Changes in N-glycan types were confirmed by lectin binding assays in both cell lines, and the rescued cell line, HuNB(-/+MGAT2). Western blotting of cells heterologously expressing a voltage-gated K+ channel (Kv3.1b) showed that some hybrid N-glycans of Kv3.1b could be processed to complex type in HuNB(-/+MGAT2) cells. In comparing HuNB and HuNB(-MGAT2) cells, decreased complex N-glycans reduced anchorage-independent cell growth, cell proliferation, and cell invasiveness, while they enhanced cell-cell interactions. Cell proliferation, invasiveness and adhesion of the HuNB(-/+MGAT2) cells were more like the HuNB than HuNB(-MGAT2). Western blotting revealed lower protein levels of MMP-2, EGFR and Gab2 in glycosylation mutant cells relative to parental cells. Gelatin zymography demonstrated that decreased MMP-2 protein activity was related to lowered MMP-2 protein levels. Thus, our results support that decreased complex type N-glycans suppress cell proliferation and cell invasiveness in both NB cell lines via remodeling ECM. MDPI 2020-04-04 /pmc/articles/PMC7236022/ /pubmed/32260356 http://dx.doi.org/10.3390/biology9040071 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hall, M. Kristen
Whitman, Austin A.
Weidner, Douglas A.
Schwalbe, Ruth A.
Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title_full Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title_fullStr Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title_full_unstemmed Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title_short Knockdown of N-Acetylglucosaminyltransferase-II Reduces Matrix Metalloproteinase 2 Activity and Suppresses Tumorigenicity in Neuroblastoma Cell Line
title_sort knockdown of n-acetylglucosaminyltransferase-ii reduces matrix metalloproteinase 2 activity and suppresses tumorigenicity in neuroblastoma cell line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7236022/
https://www.ncbi.nlm.nih.gov/pubmed/32260356
http://dx.doi.org/10.3390/biology9040071
work_keys_str_mv AT hallmkristen knockdownofnacetylglucosaminyltransferaseiireducesmatrixmetalloproteinase2activityandsuppressestumorigenicityinneuroblastomacellline
AT whitmanaustina knockdownofnacetylglucosaminyltransferaseiireducesmatrixmetalloproteinase2activityandsuppressestumorigenicityinneuroblastomacellline
AT weidnerdouglasa knockdownofnacetylglucosaminyltransferaseiireducesmatrixmetalloproteinase2activityandsuppressestumorigenicityinneuroblastomacellline
AT schwalberutha knockdownofnacetylglucosaminyltransferaseiireducesmatrixmetalloproteinase2activityandsuppressestumorigenicityinneuroblastomacellline