Cancer Res Treat.  2012 Sep;44(3):202-209.

Epidermal Growth Factor Receptor: Is It a Feasible Target for the Treatment of Osteosarcoma?

• Lee JA
• Ko Y
• Kim DH
• Lim JS
• Kong CB
• Cho WH
• Jeon DG
• Lee SY
• Koh JS

Abstract

PURPOSE
Features of epidermal growth factor receptor (EGFR) expression in osteosarcoma and in vitro efficacies of EGFR inhibitors against osteosarcoma cells were evaluated.
MATERIALS AND METHODS
Thirty biopsy samples of osteosarcoma patients were retrospectively analyzed for EGFR protein expression by immunohistochemistry. Relationships between EGFR expression and clinicopathologic characteristics and treatment outcomes were evaluated. Four osteosarcoma cell lines were analyzed for EGFR and p-EGFR expression by western blotting. Efficacies of gefitinib and BIBW2992 on osteosarcoma cells were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Tyrosine kinase domains in exons 18 to 21 were sequenced and gene expression analyses of EGFR and PTEN were performed in four osteosarcoma cell lines.
RESULTS
EGFR protein was expressed in 27 (90%) samples (6 low, 12 intermediate, 9 high) and in three cell lines. Intermediate or high staining for EGFR was related to a tumor volume<150 mL (p<0.001) and histologic subtype other than osteoblastic type (p=0.03). However, EGFR expression was not associated with histologic response to preoperative chemotherapy or survival. Gefitinib and BIBW 2992 did not have any significant inhibitory effect on cell viabilities. DNA sequencing analysis revealed three osteosarcoma cell lines have single base changes at codon 2361 of exon 20 (G to A), without affecting translation results. Furthermore, no mutation was found to be associated with constitutive EGFR activation.
CONCLUSION
In the present study, gefitinib and BIBW2992 were not effective against osteosarcoma cells. However, as osteosarcoma cells express EGFR, further studies are necessary to explore the potential of other therapeutic agents targeting EGFR.

Keyword

Epidermal growth factor receptor; Osteosarcoma; Treatment

MeSH Terms

Biopsy
Blotting, Western
Cell Line
Cell Survival
Codon
Epidermal Growth Factor
Exons
Gene Expression
Humans
Immunohistochemistry
Osteoblasts
Osteosarcoma
Protein-Tyrosine Kinases
Quinazolines
Receptor, Epidermal Growth Factor
Retrospective Studies
Sequence Analysis, DNA
Tetrazolium Salts
Thiazoles
Codon
Epidermal Growth Factor
Protein-Tyrosine Kinases
Quinazolines
Receptor, Epidermal Growth Factor
Tetrazolium Salts
Thiazoles

Figure

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  Fig. 1 Immunohistochemical staining for epidermal growth factor receptor protein in initial biopsy samples from patients with osteosarcoma. (A) Negative. (B) Low. (C) Intermediate. (D) High (A-D, ×400).

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  Fig. 2 Epidermal growth factor receptor protein expression and survival of patients with localized osteosarcoma.

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  Fig. 3 Epidermal growth factor receptor (EGFR) and p-EGFR expression of osteosarcoma cell lines in basal culture conditions.

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  Fig. 4 Effect of gefitinib and BIBW2992 on viability of osteosarcoma cells.

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