Epidermal growth factor receptor expression in squamous cell lung carcinomas: an immunohistochemical and gene analysis in formalin-fixed, paraffin-embedded material.
Epidermal growth factor (EGF) and its receptor (EGFr) constitute an important and well-characterized mitogenic system in various ectodermal tissues. We evaluated the expression of EGFr and examined possible EGFr gene alterations in 18 formalin-fixed, paraffin-embedded squamous cell lung carcinomas (SCLC) by an immunohistochemical assay, Southern blotting and differential polymerase chain reaction (DPCR). The immunohistochemical study employing the F4 and EGF-R1 monoclonal antibodies, directed against the intra- and extra-cellular portion of the receptor respectively, showed EGFr over-expression in 89% of the SCLC cases examined. All cases showed positive immunostaining for both antibodies, thus excluding the possibility of truncated receptors. In addition, analysis of the EGFr gene was carried out by Southern blotting and DPCR on paraffin extracted DNA from the same carcinoma cases. We found amplification of the EGFr gene in 5/18 (27%) SCLCs. All 5 positive cases showed EGFr over-expression, suggesting a possible correlation between the presence of EGFr gene amplification and over-expression of receptor protein. No correlation was observed among EGFr staining, EGFr gene amplification and differentiation of carcinomas. In addition, Southern blot analysis with HER-A2, a probe which hybridizes a sequence of the receptor's intracellular domain, revealed three novel EcoRI restriction fragment patterns. We suggest that these patterns correspond to EcoRI polymorphic sites of the receptor's tyrosine kinase domain.
Gorgoulis
V
V
Department of Pathology, Red Cross Hospital, Athens, Greece.
Giatromanolaki
A
A
Karameris
A
A
Tsatsanis
C
C
Aninos
D
D
Ozanne
B
B
Veslemes
M
M
Jordanoglou
J
J
Trigidou
R
R
Papastamatiou
H
H
D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000737:chemistry*; D004273:DNA, Neoplasm / Q000235:genetics*; D066246:ErbB Receptors / Q000032:analysis* / Q000235:genetics; D005557:Formaldehyde; D005784:Gene Amplification; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry*; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D016707:Tissue Fixation
D004273:DNA, Neoplasm; D005557:Formaldehyde; D066246:ErbB Receptors