Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma

Byeong-Chool Moon; Se-Jin Han; Dongjun Jeong; Kyung-Wook Kim

doi:10.5125/jkaoms.2011.37.5.396

Journal List > J Korean Assoc Oral Maxillofac Surg > v.37(5) > 1032494

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Moon, Han, Jeong, and Kim: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma

Original Article

J Korean Assoc Oral Maxillofac Surg 2011;37(5):396-402.

Published online: 10 January 2011

DOI: https://doi.org/10.5125/jkaoms.2011.37.5.396

Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma

Byeong-Chool Moon¹, Se-Jin Han¹, Dongjun Jeong², Kyung-Wook Kim¹

¹Department of Oral and Maxillofacial Surgery, School of Dentistry, Dankook University, Cheonan, Korea

²Department of Pathology, School of Medicine, Soonchunhyang University, Cheonan, Korea

Kyung-Wook Kim Department of Oral and Maxillofacial Surgery, Dental Hospital, Dankook University San 7-1, Sinbu-dong, Choenan, 330-716, Korea TEL: +82-41-550-1994 FAX: +82-41-551-8988 E-mail: kkwoms@dku.edu

Received 6 May 2011 Revised 30 September 2011 Accepted 29 September 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Introduction

Epidermal growth factor is a single-chain polypeptide consisting of 53 amino acids with potent mitogenic activity that stimulates the proliferation of a range of normal and neoplastic cells through an interaction with its specific receptor (epidermal growth factor receptor, EGFR). This interaction plays a key role in tumor progression including the induction of tumor cell proliferation. An increased EGFR copy number have been associated with a favorable response to EGFR tyrosine kinase inhibitors therapy. In contrast, K-ras mutations tend to predict a poor response to such therapy. The aim of this study was to determine the correlation between the clinicopathological factors and the upregulation of EGFR expression and K-ras mutations in oral squamous cell carcinoma.

Materials and Methods

This study examined the immunohistochemical staining of EGFR, K-ras mutation detection with peptide nucleic acid (PNA)-based real-time polymerase chain reaction (PCR) clamping in 20 specimens from 20 patients with oral squamous cell carcinoma.

Results

1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, a high level of EGFR staining was observed. The correlation between immunohistochemical EGFR expression and histological differentiation, as well as the tumor size of the specimens was significant (Pearson correlation analysis, significance [r] >0.5, P<0.05). 2. In PNA-based real-time PCR clamping analysis, a K-ras mutation was not detected in all specimens.

Conclusion

These findings suggest that the upregulation of the EGFR may play a role in the progression and invasion of oral squamous cell carcinoma that is, independent of a K-ras mutation.

Keywords: Squamous cell carcinoma, Epidermal growth factor receptor, ras Proteins

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Fig. 1.

Real-time PCR cyclining condition. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 2.

The PNA probe which is complementary to the wild type K-ras binds to the K-ras gene completely and the PCR is inhibited, while on mutant K-ras gene, the PNA probe binds to the mutan K-ras gene imcompletely and PCR is not inhibited resulting in amplicon. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 3.

Negative immunohistochemical staining for EGFR of well differentiated oral squamous cell carcinoma (anti-EGFR Immunoperoxidase, ×100). EGFR, epidermal growth factor receptor. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 4.

Mild immunohistochemical staining for EGFR of moderate differentiated oral squamous cell carcinoma (anti-EGFR Immunoperoxidase. ×100). EGFR, epidermal growth factor receptor. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 5.

Moderate immunohistochemical staining for EGFR of poor differentiated oral squamous cell carcinoma (anti-EGFR Immunoperoxidase, ×100). EGFR, epidermal growth factor receptor. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 6.

Severe immunohistochemical staining for EGFR of poor differentiated and invasive oral squamous cell carcinoma (anti-EGFR Immunoperoxidase, ×100). EGFR, epidermal growth factor receptor. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Fig. 7.

K-ras mutation detection with PNA-based real-time PCR clamping. Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

Table 1.

The correlation between immunohistochemical EGFR expression and clinical and pathological factors

Variable		Case (n)	EGFR staining pattern		r
Variable		Case (n)	Low-staining (n=)	High-staining (n=)	P
Sex	Male	11	8	3	−0.071
Sex	Female	9	7	2	0.396
Age	60≤	5	4	1	0.188
Age	60>	15	11	4	0.351
Histological differentiation	Well	13	13	0	0.697^*
	Moderate	3	2	1	0.021^*
	Poor	4	0	4
Tumor size	T1	0	0	0
	T2	11	10	1	0.599^*
	T3	7	5	2	0.032^*
	T4	2	0	2
Nodal status	N(−)	11	8	3	0.441
Nodal status	N(+)	9	7	2	0.335
Metastasis	M(−)	17	13	4	0.394
Metastasis	M(+)	3	2	1	0.239
TNM stage	I	0	0	0
	II	8	6	2	0.419
	III	6	5	1	0.072
	IV	6	4	2

n, number of patients; r, correlation coefficient; EGFR, epidermal growth factor receptor; TNM, tumor-node-metastasis

^* Pearson correlation analysis, significance [r]>0.5, P<0.05

Byeong-Chool Moon et al: Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma. J Korean Assoc Oral Maxillofac Surg 2011

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