Overexpression of bHLH domain of HIF-1 failed to inhibit the HIF-1 transcriptional activity in hypoxia.
Arnt
Basic helix-loop helix
Cancer
DNA binding
Gene therapy
HIF-1
Hypoxia
Hypoxia inducible factor
PAS
bHLH
Journal
Biological research
ISSN: 0717-6287
Titre abrégé: Biol Res
Pays: England
ID NLM: 9308271
Informations de publication
Date de publication:
05 Jun 2020
05 Jun 2020
Historique:
received:
05
12
2019
accepted:
18
05
2020
entrez:
7
6
2020
pubmed:
7
6
2020
medline:
17
6
2020
Statut:
epublish
Résumé
Hypoxia inducible factor-1 (HIF-1) is considered as the most activated transcriptional factor in response to low oxygen level or hypoxia. HIF-1 binds the hypoxia response element (HRE) sequence in the promoter of different genes, mainly through the bHLH domain and activates the transcription of genes, especially those involved in angiogenesis and EMT. Considering the critical role of bHLH in binding HIF-1 to the HRE sequence, we hypothesized that bHLH could be a promising candidate to be targeted in hypoxia condition. We inserted an inhibitory bHLH (ibHLH) domain in a pIRES2-EGFP vector and transfected HEK293T cells with either the control vector or the designed construct. The ibHLH domain consisted of bHLH domains of both HIF-1a and Arnt, capable of competing with HIF-1 in binding to HRE sequences. The transfected cells were then treated with 200 µM of cobalt chloride (CoCl Hypoxia was successfully induced in the HEK293T cell line as the gene expression of VEGF, vimentin, and β-catenin were significantly increased after treatment of untransfected HEK293T cells with 200 µM CoCl bHLH has been reported to be an important domain involved in the DNA binding activity of HIF. However, we found that targeting this domain is not sufficient to inhibit the endogenous HIF-1 transcriptional activity. Further studies about the function of critical domains of HIF-1 are necessary for developing a specific HIF-1 inhibitor.
Sections du résumé
BACKGROUND
BACKGROUND
Hypoxia inducible factor-1 (HIF-1) is considered as the most activated transcriptional factor in response to low oxygen level or hypoxia. HIF-1 binds the hypoxia response element (HRE) sequence in the promoter of different genes, mainly through the bHLH domain and activates the transcription of genes, especially those involved in angiogenesis and EMT. Considering the critical role of bHLH in binding HIF-1 to the HRE sequence, we hypothesized that bHLH could be a promising candidate to be targeted in hypoxia condition.
METHODS
METHODS
We inserted an inhibitory bHLH (ibHLH) domain in a pIRES2-EGFP vector and transfected HEK293T cells with either the control vector or the designed construct. The ibHLH domain consisted of bHLH domains of both HIF-1a and Arnt, capable of competing with HIF-1 in binding to HRE sequences. The transfected cells were then treated with 200 µM of cobalt chloride (CoCl
RESULTS
RESULTS
Hypoxia was successfully induced in the HEK293T cell line as the gene expression of VEGF, vimentin, and β-catenin were significantly increased after treatment of untransfected HEK293T cells with 200 µM CoCl
CONCLUSION
CONCLUSIONS
bHLH has been reported to be an important domain involved in the DNA binding activity of HIF. However, we found that targeting this domain is not sufficient to inhibit the endogenous HIF-1 transcriptional activity. Further studies about the function of critical domains of HIF-1 are necessary for developing a specific HIF-1 inhibitor.
Identifiants
pubmed: 32503642
doi: 10.1186/s40659-020-00293-4
pii: 10.1186/s40659-020-00293-4
pmc: PMC7275393
doi:
Substances chimiques
Basic Helix-Loop-Helix Transcription Factors
0
Hypoxia-Inducible Factor 1
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
25Subventions
Organisme : Iran University of Medical Sciences
ID : 31242
Références
Cell Cycle. 2004 Feb;3(2):164-7
pubmed: 14712082
Oncol Lett. 2018 Jan;15(1):1119-1124
pubmed: 29391899
Mediators Inflamm. 2015;2015:584758
pubmed: 26491231
Gynecol Endocrinol. 2016;32(4):323-8
pubmed: 26654708
J Med Chem. 2013 Feb 28;56(4):1739-47
pubmed: 23363003
Mol Cell Biol. 1995 Feb;15(2):756-65
pubmed: 7823943
Sci Rep. 2015 Dec 09;5:17851
pubmed: 26647819
Mol Cell Biol. 1998 Jul;18(7):4079-88
pubmed: 9632792
Sci Rep. 2017 Nov 21;7(1):15918
pubmed: 29162839
Nature. 1993 May 6;363(6424):38-45
pubmed: 8479534
J Biomol Struct Dyn. 2000 Oct;18(2):169-79
pubmed: 11089639
Genes Cancer. 2011 Dec;2(12):1117-33
pubmed: 22866203
Recent Pat Anticancer Drug Discov. 2019;14(2):158-169
pubmed: 31131753
Cancer Metastasis Rev. 2007 Jun;26(2):225-39
pubmed: 17440684
Exp Ther Med. 2017 Aug;14(2):1825-1831
pubmed: 28810656
Oral Oncol. 2018 May;80:23-32
pubmed: 29706185
Sci Rep. 2017 Jan 11;7:40446
pubmed: 28074862
Cell Death Differ. 2008 Apr;15(4):642-9
pubmed: 18202699
Nature. 2015 Aug 20;524(7565):303-8
pubmed: 26245371
Mol Cell Biol. 1996 Oct;16(10):5865-75
pubmed: 8927054
Oncologist. 2004;9 Suppl 5:31-40
pubmed: 15591420
J Biol Chem. 2002 Sep 6;277(36):32405-8
pubmed: 12119283
J Vis Exp. 2011 Aug 12;(54):
pubmed: 21860378
Int J Oncol. 2016 Jan;48(1):271-80
pubmed: 26648580
J Biol Chem. 1996 Feb 2;271(5):2683-8
pubmed: 8576241
Acta Pharm Sin B. 2015 Sep;5(5):378-89
pubmed: 26579469
Einstein (Sao Paulo). 2017 Jul-Sep;15(3):369-375
pubmed: 29091160
J Biol Chem. 2003 May 2;278(18):15911-6
pubmed: 12606543
ChemMedChem. 2016 Apr 19;11(8):773-86
pubmed: 26997519
Virchows Arch. 2010 Jul;457(1):53-61
pubmed: 20526721
Nat Cell Biol. 2007 Feb;9(2):210-7
pubmed: 17220880
Proc Natl Acad Sci U S A. 2004 Nov 30;101(48):16768-73
pubmed: 15556999
Hypoxia (Auckl). 2015 Dec 11;3:83-92
pubmed: 27774485
Cell Death Dis. 2019 Dec 9;10(12):939
pubmed: 31819036
Eur J Biochem. 1997 Jun 1;246(2):486-95
pubmed: 9208942
Mol Cell Biol. 1999 Aug;19(8):5811-22
pubmed: 10409767
Stem Cells. 2013 Sep;31(9):1902-9
pubmed: 23744828
Sci Rep. 2015 Jul 15;5:12410
pubmed: 26174737
Nat Protoc. 2007;2(8):1849-61
pubmed: 17703195
Sci STKE. 2005 Oct 18;2005(306):re12
pubmed: 16234508
Cancer Res. 2005 Oct 1;65(19):9047-55
pubmed: 16204079
J Intern Med. 2013 Aug;274(2):105-12
pubmed: 23844914
Trends Biochem Sci. 2008 Nov;33(11):526-34
pubmed: 18809331
Front Cell Dev Biol. 2016 Feb 23;4:11
pubmed: 26942179
Trends Cancer. 2016 Dec;2(12):758-770
pubmed: 28741521
Cell Cycle. 2006 Aug;5(16):1847-53
pubmed: 16861921
Nature. 2001 Nov 29;414(6863):550-4
pubmed: 11734856