Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma

Hiromitsu Hatakeyama, Haixia Cheng, Pamela Wirth, Ashley Counsell, Samuel R. Marcrom, Carey Burton Wood, Paula R. Pohlmann, Jill Gilbert, Barbara Murphy, Wendell G. Yarbrough, Deric L. Wheeler, Paul M. Harari, Yan Guo, Yu Shyr, Robbert J. Slebos, Christine H. Chung

Research output: Contribution to journalArticle

Abstract

Background: We hypothesized that chronic inhibition of epidermal growth factor receptor (EGFR) by cetuximab, a monoclonal anti-EGFR antibody, induces up-regulation of its ligands resulting in resistance and that microRNAs (miRs) play an important role in the ligand regulation in head and neck squamous cell carcinoma (HNSCC). Methodology/Principal Findings: Genome-wide changes in gene and miR expression were determined in cetuximabsensitive cell line, SCC1, and its resistant derivative 1Cc8 using DNA microarrays and RT-PCR. The effects of differentially expressed EGFR ligands and miRs were examined by MTS, colony formation, ELISA, and western blot assays. Heparinbinding EGF-like growth factor (HB-EGF) and its regulator, miR-212, were differentially expressed with statistical significance when SCC1 and 1Cc8 were compared for gene and miR expression. Stimulation with HB-EGF induced cetuximab resistance in sensitive cell lines. Inhibition of HB-EGF and the addition of miR-212 mimic induced cetuximab sensitivity in resistant cell lines. MicroRNA-212 and HB-EGF expression were inversely correlated in an additional 33 HNSCC and keratinocyte cell lines. Six tumors and 46 plasma samples from HNSCC patients were examined for HB-EGF levels. HB-EGF plasma levels were lower in newly diagnosed HNSCC patients when compared to patients with recurrent disease. Conclusions/Significance: Increased expression of HB-EGF due to down-regulation of miR-212 is a possible mechanism of cetuximab resistance. The combination of EGFR ligand inhibitors or miR modulators with cetuximab may improve the clinical outcome of cetuximab therapy in HNSCC.

Original languageEnglish (US)
Article numbere12702
Pages (from-to)1-13
Number of pages13
JournalPLoS One
Volume5
Issue number9
DOIs
StatePublished - 2010

Fingerprint

squamous cell carcinoma
heparin
Epidermal Growth Factor
growth factors
neck
Intercellular Signaling Peptides and Proteins
Epidermal Growth Factor Receptor
MicroRNAs
microRNA
Cells
cell lines
Ligands
Cell Line
Genes
Plasmas
Gene Expression
induced resistance
keratinocytes
Microarrays
Epithelial Cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Hatakeyama, H., Cheng, H., Wirth, P., Counsell, A., Marcrom, S. R., Wood, C. B., ... Chung, C. H. (2010). Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma. PLoS One, 5(9), 1-13. [e12702]. https://doi.org/10.1371/journal.pone.0012702

Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma. / Hatakeyama, Hiromitsu; Cheng, Haixia; Wirth, Pamela; Counsell, Ashley; Marcrom, Samuel R.; Wood, Carey Burton; Pohlmann, Paula R.; Gilbert, Jill; Murphy, Barbara; Yarbrough, Wendell G.; Wheeler, Deric L.; Harari, Paul M.; Guo, Yan; Shyr, Yu; Slebos, Robbert J.; Chung, Christine H.

In: PLoS One, Vol. 5, No. 9, e12702, 2010, p. 1-13.

Research output: Contribution to journalArticle

Hatakeyama, H, Cheng, H, Wirth, P, Counsell, A, Marcrom, SR, Wood, CB, Pohlmann, PR, Gilbert, J, Murphy, B, Yarbrough, WG, Wheeler, DL, Harari, PM, Guo, Y, Shyr, Y, Slebos, RJ & Chung, CH 2010, 'Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma', PLoS One, vol. 5, no. 9, e12702, pp. 1-13. https://doi.org/10.1371/journal.pone.0012702
Hatakeyama, Hiromitsu ; Cheng, Haixia ; Wirth, Pamela ; Counsell, Ashley ; Marcrom, Samuel R. ; Wood, Carey Burton ; Pohlmann, Paula R. ; Gilbert, Jill ; Murphy, Barbara ; Yarbrough, Wendell G. ; Wheeler, Deric L. ; Harari, Paul M. ; Guo, Yan ; Shyr, Yu ; Slebos, Robbert J. ; Chung, Christine H. / Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma. In: PLoS One. 2010 ; Vol. 5, No. 9. pp. 1-13.
@article{84ddc2e2cafa46b88665ead0da0c19aa,
title = "Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma",
abstract = "Background: We hypothesized that chronic inhibition of epidermal growth factor receptor (EGFR) by cetuximab, a monoclonal anti-EGFR antibody, induces up-regulation of its ligands resulting in resistance and that microRNAs (miRs) play an important role in the ligand regulation in head and neck squamous cell carcinoma (HNSCC). Methodology/Principal Findings: Genome-wide changes in gene and miR expression were determined in cetuximabsensitive cell line, SCC1, and its resistant derivative 1Cc8 using DNA microarrays and RT-PCR. The effects of differentially expressed EGFR ligands and miRs were examined by MTS, colony formation, ELISA, and western blot assays. Heparinbinding EGF-like growth factor (HB-EGF) and its regulator, miR-212, were differentially expressed with statistical significance when SCC1 and 1Cc8 were compared for gene and miR expression. Stimulation with HB-EGF induced cetuximab resistance in sensitive cell lines. Inhibition of HB-EGF and the addition of miR-212 mimic induced cetuximab sensitivity in resistant cell lines. MicroRNA-212 and HB-EGF expression were inversely correlated in an additional 33 HNSCC and keratinocyte cell lines. Six tumors and 46 plasma samples from HNSCC patients were examined for HB-EGF levels. HB-EGF plasma levels were lower in newly diagnosed HNSCC patients when compared to patients with recurrent disease. Conclusions/Significance: Increased expression of HB-EGF due to down-regulation of miR-212 is a possible mechanism of cetuximab resistance. The combination of EGFR ligand inhibitors or miR modulators with cetuximab may improve the clinical outcome of cetuximab therapy in HNSCC.",
author = "Hiromitsu Hatakeyama and Haixia Cheng and Pamela Wirth and Ashley Counsell and Marcrom, {Samuel R.} and Wood, {Carey Burton} and Pohlmann, {Paula R.} and Jill Gilbert and Barbara Murphy and Yarbrough, {Wendell G.} and Wheeler, {Deric L.} and Harari, {Paul M.} and Yan Guo and Yu Shyr and Slebos, {Robbert J.} and Chung, {Christine H.}",
year = "2010",
doi = "10.1371/journal.pone.0012702",
language = "English (US)",
volume = "5",
pages = "1--13",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "9",

}

TY - JOUR

T1 - Regulation of heparin-binding EGF-like growth factor by miR-212 and acquired cetuximab-resistance in head and neck squamous cell carcinoma

AU - Hatakeyama, Hiromitsu

AU - Cheng, Haixia

AU - Wirth, Pamela

AU - Counsell, Ashley

AU - Marcrom, Samuel R.

AU - Wood, Carey Burton

AU - Pohlmann, Paula R.

AU - Gilbert, Jill

AU - Murphy, Barbara

AU - Yarbrough, Wendell G.

AU - Wheeler, Deric L.

AU - Harari, Paul M.

AU - Guo, Yan

AU - Shyr, Yu

AU - Slebos, Robbert J.

AU - Chung, Christine H.

PY - 2010

Y1 - 2010

N2 - Background: We hypothesized that chronic inhibition of epidermal growth factor receptor (EGFR) by cetuximab, a monoclonal anti-EGFR antibody, induces up-regulation of its ligands resulting in resistance and that microRNAs (miRs) play an important role in the ligand regulation in head and neck squamous cell carcinoma (HNSCC). Methodology/Principal Findings: Genome-wide changes in gene and miR expression were determined in cetuximabsensitive cell line, SCC1, and its resistant derivative 1Cc8 using DNA microarrays and RT-PCR. The effects of differentially expressed EGFR ligands and miRs were examined by MTS, colony formation, ELISA, and western blot assays. Heparinbinding EGF-like growth factor (HB-EGF) and its regulator, miR-212, were differentially expressed with statistical significance when SCC1 and 1Cc8 were compared for gene and miR expression. Stimulation with HB-EGF induced cetuximab resistance in sensitive cell lines. Inhibition of HB-EGF and the addition of miR-212 mimic induced cetuximab sensitivity in resistant cell lines. MicroRNA-212 and HB-EGF expression were inversely correlated in an additional 33 HNSCC and keratinocyte cell lines. Six tumors and 46 plasma samples from HNSCC patients were examined for HB-EGF levels. HB-EGF plasma levels were lower in newly diagnosed HNSCC patients when compared to patients with recurrent disease. Conclusions/Significance: Increased expression of HB-EGF due to down-regulation of miR-212 is a possible mechanism of cetuximab resistance. The combination of EGFR ligand inhibitors or miR modulators with cetuximab may improve the clinical outcome of cetuximab therapy in HNSCC.

AB - Background: We hypothesized that chronic inhibition of epidermal growth factor receptor (EGFR) by cetuximab, a monoclonal anti-EGFR antibody, induces up-regulation of its ligands resulting in resistance and that microRNAs (miRs) play an important role in the ligand regulation in head and neck squamous cell carcinoma (HNSCC). Methodology/Principal Findings: Genome-wide changes in gene and miR expression were determined in cetuximabsensitive cell line, SCC1, and its resistant derivative 1Cc8 using DNA microarrays and RT-PCR. The effects of differentially expressed EGFR ligands and miRs were examined by MTS, colony formation, ELISA, and western blot assays. Heparinbinding EGF-like growth factor (HB-EGF) and its regulator, miR-212, were differentially expressed with statistical significance when SCC1 and 1Cc8 were compared for gene and miR expression. Stimulation with HB-EGF induced cetuximab resistance in sensitive cell lines. Inhibition of HB-EGF and the addition of miR-212 mimic induced cetuximab sensitivity in resistant cell lines. MicroRNA-212 and HB-EGF expression were inversely correlated in an additional 33 HNSCC and keratinocyte cell lines. Six tumors and 46 plasma samples from HNSCC patients were examined for HB-EGF levels. HB-EGF plasma levels were lower in newly diagnosed HNSCC patients when compared to patients with recurrent disease. Conclusions/Significance: Increased expression of HB-EGF due to down-regulation of miR-212 is a possible mechanism of cetuximab resistance. The combination of EGFR ligand inhibitors or miR modulators with cetuximab may improve the clinical outcome of cetuximab therapy in HNSCC.

UR - http://www.scopus.com/inward/record.url?scp=77958520234&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77958520234&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0012702

DO - 10.1371/journal.pone.0012702

M3 - Article

C2 - 20856931

AN - SCOPUS:77958520234

VL - 5

SP - 1

EP - 13

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 9

M1 - e12702

ER -