Reference MicroRNAs for RT-qPCR Assays in Cervical Cancer Patients and Their Application to Studies of HPV16 and Hypoxia Biomarkers.
Journal
Translational oncology
ISSN: 1936-5233
Titre abrégé: Transl Oncol
Pays: United States
ID NLM: 101472619
Informations de publication
Date de publication:
Mar 2019
Mar 2019
Historique:
received:
25
10
2018
revised:
19
12
2018
accepted:
20
12
2018
pubmed:
21
1
2019
medline:
21
1
2019
entrez:
21
1
2019
Statut:
ppublish
Résumé
MicroRNA (miRNA) expressions in tumor biopsies have shown potential as biomarkers in cervical cancer, but suitable reference RNAs for normalization of reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays in patient cohorts with different clinicopathological characteristics are not available. We aimed to identify the optimal reference miRNAs and apply these to investigate the potential of miR-9-5p as human papilloma virus (HPV) 16 biomarker and miR-210-3p as hypoxia biomarker in cervical cancer. Candidate reference miRNAs were preselected in sequencing data of 90 patients and ranked in a stability analysis by RefFinder. A selection of the most stable miRNAs was evaluated by geNorm and NormFinder analyses of RT-qPCR data of 29 patients. U6 small nuclear RNA (RNU6) was also included in the evaluation. MiR-9-5p and miR-210-3p expression was assessed by RT-qPCR in 45 and 65 patients, respectively. Nine candidates were preselected in the sequencing data after excluding those associated with clinical markers, HPV type, hypoxia status, suboptimal expression levels, and low stability. In RT-qPCR assays, the combination of miR-151-5p, miR-152-3p, and miR-423-3p was identified as the most stable normalization factor across clinical markers, HPV type, and hypoxia status. RNU6 showed poor stability. By applying the optimal reference miRNAs, higher miR-9-5p expression in HPV16- than HPV18-positive tumors and higher miR-210-3p expression in more hypoxic than less hypoxic tumors were found in accordance with the sequencing data. MiR-210-3p was associated with poor outcome by both sequencing and RT-qPCR assays. In conclusion, miR-151-5p, miR-152-3p, and miR-423-3p are suitable reference miRNAs in cervical cancer. MiR-9-5p and miR-210-3p are promising HPV16 and hypoxia biomarkers, respectively.
Identifiants
pubmed: 30660934
pii: S1936-5233(18)30544-8
doi: 10.1016/j.tranon.2018.12.010
pmc: PMC6349320
pii:
doi:
Types de publication
Journal Article
Langues
eng
Pagination
576-584Informations de copyright
Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.
Références
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Genome Biol. 2002 Jun 18;3(7):RESEARCH0034
pubmed: 12184808
Biotechnol Lett. 2004 Mar;26(6):509-15
pubmed: 15127793
Cancer Res. 2004 Aug 1;64(15):5245-50
pubmed: 15289330
Nucleic Acids Res. 2006 Jan 1;34(Database issue):D140-4
pubmed: 16381832
BMC Mol Biol. 2006 Oct 06;7:33
pubmed: 17026756
RNA. 2008 May;14(5):844-52
pubmed: 18375788
Clin Cancer Res. 2008 May 1;14(9):2535-42
pubmed: 18451214
Cancer Inform. 2007 Feb 04;3:11-7
pubmed: 19455231
Cancer Res. 2010 Feb 15;70(4):1441-8
pubmed: 20124485
BMC Genomics. 2011 Mar 21;12:156
pubmed: 21418615
Exp Mol Med. 2011 Jun 30;43(6):358-66
pubmed: 21519184
Anal Biochem. 2011 Oct 15;417(2):233-41
pubmed: 21741950
Nucleic Acids Res. 2012 Jan;40(1):37-52
pubmed: 21911355
Plant Mol Biol. 2012 Jan 31;:null
pubmed: 22290409
J Pathol. 2013 May;230(1):59-69
pubmed: 23335387
Anticancer Res. 2013 Jun;33(6):2561-7
pubmed: 23749909
Antioxid Redox Signal. 2014 Sep 10;21(8):1220-38
pubmed: 24111776
BMC Res Notes. 2014 Mar 07;7:129
pubmed: 24606728
Methods Mol Biol. 2014;1160:19-26
pubmed: 24740218
Gynecol Oncol. 2014 Jul;134(1):129-37
pubmed: 24793973
Oncotarget. 2014 Nov 30;5(22):11620-30
pubmed: 25344913
PLoS One. 2014 Nov 03;9(11):e111021
pubmed: 25365304
FEBS Lett. 2015 Mar 12;589(6):702-9
pubmed: 25666710
Tumour Biol. 2015 Aug;36(8):5825-30
pubmed: 25712373
PLoS One. 2015 Mar 24;10(3):e0120905
pubmed: 25803820
PLoS One. 2015 Mar 31;10(3):e0122515
pubmed: 25825906
PLoS One. 2015 Apr 16;10(4):e0123946
pubmed: 25880806
Int J Clin Exp Pathol. 2015 May 01;8(5):5542-8
pubmed: 26191262
Clin Chem. 2015 Nov;61(11):1333-42
pubmed: 26408530
Eur Rev Med Pharmacol Sci. 2015 Nov;19(21):4081-5
pubmed: 26592830
Oncotarget. 2016 Apr 12;7(15):19666-79
pubmed: 26910911
Clin Cancer Res. 2016 Aug 15;22(16):4067-76
pubmed: 27012812
Int J Gynecol Cancer. 2016 Jun;26(5):817-24
pubmed: 27206216
PLoS One. 2016 May 31;11(5):e0156259
pubmed: 27244197
J Obstet Gynaecol Res. 2016 Sep;42(9):1168-79
pubmed: 27279231
QJM. 2017 Jan;110(1):11-15
pubmed: 27345415
Biomed Pharmacother. 2016 Oct;83:64-69
pubmed: 27470551
Future Oncol. 2017 Apr;13(8):743-753
pubmed: 27806630
Int J Gynecol Cancer. 2017 Feb;27(2):339-343
pubmed: 27870701
Sci Rep. 2017 Jul 17;7(1):5624
pubmed: 28717180
Clin Transl Imaging. 2017;5(4):373-388
pubmed: 28804704
J Mol Diagn. 2017 Sep;19(5):625-637
pubmed: 28826607
Gene. 2018 Feb 5;642:256-260
pubmed: 29154871