Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.

The stabilisation of HIF-α is central to the transcriptional response of animals to hypoxia, regulating the expression of hundreds of genes including those involved in angiogenesis, metabolism and metastasis. HIF-α is degraded under normoxic conditions by proline hydroxylation, which allows for reco...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Cameron E Snell, Helen Turley, Alan McIntyre, Demin Li, Massimo Masiero, Christopher J Schofield, Kevin C Gatter, Adrian L Harris, Francesco Pezzella
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d61ea88e516e49f18f6fe0fe2b4b0515
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:d61ea88e516e49f18f6fe0fe2b4b0515
record_format	dspace
spelling	oai:doaj.org-article:d61ea88e516e49f18f6fe0fe2b4b05152021-11-18T08:32:42ZProline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.1932-620310.1371/journal.pone.0088955https://doaj.org/article/d61ea88e516e49f18f6fe0fe2b4b05152014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24563687/?tool=EBIhttps://doaj.org/toc/1932-6203The stabilisation of HIF-α is central to the transcriptional response of animals to hypoxia, regulating the expression of hundreds of genes including those involved in angiogenesis, metabolism and metastasis. HIF-α is degraded under normoxic conditions by proline hydroxylation, which allows for recognition and ubiquitination by the von-Hippel-Lindau (VHL) E3 ligase complex. The aim of our study was to investigate the posttranslational modification of HIF-1α in tumours, to assess whether there are additional mechanisms besides reduced hydroxylation leading to stability. To this end we optimised antibodies against the proline-hydroxylated forms of HIF-1α for use in formalin fixed paraffin embedded (FFPE) immunohistochemistry to assess effects in tumour cells in vivo. We found that HIF-1α proline-hydroxylated at both VHL binding sites (Pro402 and Pro564), was present in hypoxic regions of a wide range of tumours, tumour xenografts and in moderately hypoxic cells in vitro. Staining for hydroxylated HIF-1α can identify a subset of breast cancer patients with poorer prognosis and may be a better marker than total HIF-1α levels. The expression of unhydroxylated HIF-1α positively correlates with VHL in breast cancer suggesting that VHL may be rate-limiting for HIF degradation. Our conclusions are that the degradation of proline-hydroxylated HIF-1α may be rate-limited in tumours and therefore provides new insights into mechanisms of HIF upregulation. Persistence of proline-hydroxylated HIF-1α in perinecrotic areas suggests there is adequate oxygen to support prolyl hydroxylase domain (PHD) activity and proline-hydroxylated HIF-1α may be the predominant form associated with the poorer prognosis that higher levels of HIF-1α confer.Cameron E SnellHelen TurleyAlan McIntyreDemin LiMassimo MasieroChristopher J SchofieldKevin C GatterAdrian L HarrisFrancesco PezzellaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 2, p e88955 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cameron E Snell Helen Turley Alan McIntyre Demin Li Massimo Masiero Christopher J Schofield Kevin C Gatter Adrian L Harris Francesco Pezzella Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
description	The stabilisation of HIF-α is central to the transcriptional response of animals to hypoxia, regulating the expression of hundreds of genes including those involved in angiogenesis, metabolism and metastasis. HIF-α is degraded under normoxic conditions by proline hydroxylation, which allows for recognition and ubiquitination by the von-Hippel-Lindau (VHL) E3 ligase complex. The aim of our study was to investigate the posttranslational modification of HIF-1α in tumours, to assess whether there are additional mechanisms besides reduced hydroxylation leading to stability. To this end we optimised antibodies against the proline-hydroxylated forms of HIF-1α for use in formalin fixed paraffin embedded (FFPE) immunohistochemistry to assess effects in tumour cells in vivo. We found that HIF-1α proline-hydroxylated at both VHL binding sites (Pro402 and Pro564), was present in hypoxic regions of a wide range of tumours, tumour xenografts and in moderately hypoxic cells in vitro. Staining for hydroxylated HIF-1α can identify a subset of breast cancer patients with poorer prognosis and may be a better marker than total HIF-1α levels. The expression of unhydroxylated HIF-1α positively correlates with VHL in breast cancer suggesting that VHL may be rate-limiting for HIF degradation. Our conclusions are that the degradation of proline-hydroxylated HIF-1α may be rate-limited in tumours and therefore provides new insights into mechanisms of HIF upregulation. Persistence of proline-hydroxylated HIF-1α in perinecrotic areas suggests there is adequate oxygen to support prolyl hydroxylase domain (PHD) activity and proline-hydroxylated HIF-1α may be the predominant form associated with the poorer prognosis that higher levels of HIF-1α confer.
format	article
author	Cameron E Snell Helen Turley Alan McIntyre Demin Li Massimo Masiero Christopher J Schofield Kevin C Gatter Adrian L Harris Francesco Pezzella
author_facet	Cameron E Snell Helen Turley Alan McIntyre Demin Li Massimo Masiero Christopher J Schofield Kevin C Gatter Adrian L Harris Francesco Pezzella
author_sort	Cameron E Snell
title	Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
title_short	Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
title_full	Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
title_fullStr	Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
title_full_unstemmed	Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.
title_sort	proline-hydroxylated hypoxia-inducible factor 1α (hif-1α) upregulation in human tumours.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/d61ea88e516e49f18f6fe0fe2b4b0515
work_keys_str_mv	AT cameronesnell prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT helenturley prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT alanmcintyre prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT deminli prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT massimomasiero prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT christopherjschofield prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT kevincgatter prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT adrianlharris prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours AT francescopezzella prolinehydroxylatedhypoxiainduciblefactor1ahif1aupregulationinhumantumours
_version_	1718421650611896320

Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α) upregulation in human tumours.

Ejemplares similares