Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.

Aberrant glycosylation is a common feature of many malignancies including colorectal cancers (CRCs). About 15% of CRC show the microsatellite instability (MSI) phenotype that is associated with a high frequency of biallelic frameshift mutations in the A10 coding mononucleotide microsatellite of the...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jennifer Lee, Seda Ballikaya, Kai Schönig, Claudia R Ball, Hanno Glimm, Juergen Kopitz, Johannes Gebert
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/13ae7ad23eab4a008f134de3592a642a
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:13ae7ad23eab4a008f134de3592a642a
record_format	dspace
spelling	oai:doaj.org-article:13ae7ad23eab4a008f134de3592a642a2021-11-18T07:55:41ZTransforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.1932-620310.1371/journal.pone.0057074https://doaj.org/article/13ae7ad23eab4a008f134de3592a642a2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23468914/?tool=EBIhttps://doaj.org/toc/1932-6203Aberrant glycosylation is a common feature of many malignancies including colorectal cancers (CRCs). About 15% of CRC show the microsatellite instability (MSI) phenotype that is associated with a high frequency of biallelic frameshift mutations in the A10 coding mononucleotide microsatellite of the transforming growth factor beta receptor 2 (TGFBR2) gene. If and how impaired TGFBR2 signaling in MSI CRC cells affects cell surface glycan pattern is largely unexplored. Here, we used the TGFBR2-deficient MSI colon carcinoma cell line HCT116 as a model system. Stable clones conferring doxycycline (dox)-inducible expression of a single copy wildtype TGFBR2 transgene were generated by recombinase-mediated cassette exchange (RMCE). In two independent clones, dox-inducible expression of wildtype TGFBR2 protein and reconstitution of its signaling function was shown. Metabolic labeling experiments using the tritiated sialic acid precursor N-acetyl-D-mannosamine (ManNAc) revealed a significant decline (∼30%) of its incorporation into newly synthesized sialoglycoproteins in a TGFBR2-dependent manner. In particular, we detected a significant decrease of sialylated ß1-integrin upon reconstituted TGFBR2 signaling which did not influence ß1-integrin protein turnover. Notably, TGFBR2 reconstitution did not affect the transcript levels of any of the known human sialyltransferases when examined by real-time RT- PCR analysis. These results suggest that reconstituted TGFBR2 signaling in an isogenic MSI cell line model system can modulate sialylation of cell surface proteins like ß1-integrin. Moreover, our model system will be suitable to uncover the underlying molecular mechanisms of altered MSI tumor glycobiology.Jennifer LeeSeda BallikayaKai SchönigClaudia R BallHanno GlimmJuergen KopitzJohannes GebertPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e57074 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jennifer Lee Seda Ballikaya Kai Schönig Claudia R Ball Hanno Glimm Juergen Kopitz Johannes Gebert Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
description	Aberrant glycosylation is a common feature of many malignancies including colorectal cancers (CRCs). About 15% of CRC show the microsatellite instability (MSI) phenotype that is associated with a high frequency of biallelic frameshift mutations in the A10 coding mononucleotide microsatellite of the transforming growth factor beta receptor 2 (TGFBR2) gene. If and how impaired TGFBR2 signaling in MSI CRC cells affects cell surface glycan pattern is largely unexplored. Here, we used the TGFBR2-deficient MSI colon carcinoma cell line HCT116 as a model system. Stable clones conferring doxycycline (dox)-inducible expression of a single copy wildtype TGFBR2 transgene were generated by recombinase-mediated cassette exchange (RMCE). In two independent clones, dox-inducible expression of wildtype TGFBR2 protein and reconstitution of its signaling function was shown. Metabolic labeling experiments using the tritiated sialic acid precursor N-acetyl-D-mannosamine (ManNAc) revealed a significant decline (∼30%) of its incorporation into newly synthesized sialoglycoproteins in a TGFBR2-dependent manner. In particular, we detected a significant decrease of sialylated ß1-integrin upon reconstituted TGFBR2 signaling which did not influence ß1-integrin protein turnover. Notably, TGFBR2 reconstitution did not affect the transcript levels of any of the known human sialyltransferases when examined by real-time RT- PCR analysis. These results suggest that reconstituted TGFBR2 signaling in an isogenic MSI cell line model system can modulate sialylation of cell surface proteins like ß1-integrin. Moreover, our model system will be suitable to uncover the underlying molecular mechanisms of altered MSI tumor glycobiology.
format	article
author	Jennifer Lee Seda Ballikaya Kai Schönig Claudia R Ball Hanno Glimm Juergen Kopitz Johannes Gebert
author_facet	Jennifer Lee Seda Ballikaya Kai Schönig Claudia R Ball Hanno Glimm Juergen Kopitz Johannes Gebert
author_sort	Jennifer Lee
title	Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
title_short	Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
title_full	Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
title_fullStr	Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
title_full_unstemmed	Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.
title_sort	transforming growth factor beta receptor 2 (tgfbr2) changes sialylation in the microsatellite unstable (msi) colorectal cancer cell line hct116.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/13ae7ad23eab4a008f134de3592a642a
work_keys_str_mv	AT jenniferlee transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT sedaballikaya transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT kaischonig transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT claudiarball transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT hannoglimm transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT juergenkopitz transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116 AT johannesgebert transforminggrowthfactorbetareceptor2tgfbr2changessialylationinthemicrosatelliteunstablemsicolorectalcancercelllinehct116
_version_	1718422712712429568

Transforming growth factor beta receptor 2 (TGFBR2) changes sialylation in the microsatellite unstable (MSI) Colorectal cancer cell line HCT116.

Ejemplares similares