Immunohistochemical characterizationof nitric oxide synthase activity in squamous cell carcinoma of the head and neck,☆☆,

Presented at the Annual Meeting of the American Academy of Otolaryngology–Head and Neck Surgery, New Orleans, La., Sept. 17-20, 1995.
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Abstract

This study was designed to investigate the presence of nitric oxide in human squamous cell carcinoma of the head and neck. We localized the activity of nitric oxide synthase in these tumors through immunohistochemical analysis using antibodies to L-citrulline (a byproduct of nitric oxide synthase), to inducible nitric oxide synthase, and to constitutive nitric oxide synthase. We found presence of inducible enzyme in squamous cells throughout these tumors, with the highest intensity staining occurring directly around keratin pearls. Our findings suggest that inducible nitric oxide synthase activity is present in squamous cell carcinomas of the head and neck, leading us to conclude that inducible nitric oxide synthase may play a significant role in tumor growth. (OTOLARYNGOL HEAD NECK SURG 1995;113:541-9.)

Section snippets

METHODS AND MATERIAL

In this project, 10 squamous cell carcinomas and 5 normal mucosa controls were investigated. Tumor tissue not necessary for diagnosis, margin determination, or node assessment was obtained at the time of surgical resection from 10 patients. As a control, normal oral mucosa was obtained at the time of tonsillectomy for tonsillar hypertrophy or uvulopalatoplasty for snoring from 5 patients. All tissue was obtained under approval of the patient specimen protocol of the Institutional Committee on

RESULTS

Immunoreactivity to the anticitrulline antibody and to the anti-iNOS antibody was present in squamous epithelial cells of 8 of 10 tumor specimens. Staining intensity of anticitrulline and anti-iNOS was darkest in the squamous cells directly surrounding the keratin pearls but was also present throughout the full tumor thickness (Fig. 1). Immunoreactivity to the anti-eNOS antibody was found in the vascular endothelium of 4 of 10 tumor specimens (Table 1 and Fig. 2). No obvious correlations were

DISCUSSION

From our studies, it appears that iNOS is present in squamous epithelial cells throughout squamous cell carcinoma tumors and that enzyme activity is greatest directly surrounding keratin pearls. We also demonstrated evidence of iNOS in the outermost two to three layers of normal squamous cell epithelium.

This is the first study to identify iNOS in normal squamous cell epithelium and in squamous cell carcinoma. However, NO has been previously identified in murine mammary adenocarcinoma,

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    From the Division of Otolaryngology, Head and Neck Surgery, Department of Surgery (Drs. Rosbe, Prazma, Mims, Ball, and Weissler), University of North Carolina School of Medicine; and the Department of Cell Biology and Anatomy (Dr. Petrusz), University of North Carolina.

    ☆☆

    Reprint requests: Kristina W. Rosbe, MD, Division of Otolaryngology, Head and Neck Surgery, Department of Surgery, University of North Carolina School of Medicine, 610 Burnett-Womack Building, CB -x7070, Chapel Hill, NC 27599-7070.

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