Early progression stage of malignancy as revealed by immunohistochemical demonstration of DNA instability; II, Otorhinolaryngeal border-line neoplastic lesions.

The degree of DNA-instability as revealed by the immunohistochemical staining with anti-single-stranded DNA antibody after acid hydrolysis (DNA-instability test) was used as a marker of malignancy. This was applied to benign, border-line, and malignant neoplastic lesions found in the otorhinolaryngeal regions (31 cancer, 22 leukoplakia, 10 nasal inverted papilloma, 33 salivary gland pleomorphic adenoma, and 7 Warthin's tumor cases). Proliferative activity and polarity of the proliferative cell distribution were evaluated by PCNA-immunohistochemistry, and the quantitative analyses of the number, mean size, largest size, and maximum shape-irregularity of AgNORs in a nucleus were performed for all these cases. As the results, 31 cancer (100%), 20 leukoplakia (90.1%), 10 nasal inverted papilloma (100%), and 21 pleomorphic adenoma (63.6%) cases were positively stained by the DNA-instability test diffusely or sporadically, indicating their malignancy. Reflecting the malignant character, these cases showed a remarkable increase in the PCNA-index with the loss of polarity of PCNA-positive cell distribution, and also increased number, mean and largest sizes, and maximum shape-irregularity of AgNORs. These results indicate that all nasal inverted papillomas are malignant in nature, namely, in situ carcinoma, and the majority of leukoplakia is also regarded as in situ cancer, although a certain percentage of simple hyperplasia may be included. Furthermore, the pleomorphic adenoma of the salivary gland can be regarded as an "unstable tumor" which often contains or predisposes to bear malignant subclones with occasional capsular or extracapsular invasion, reflecting the progression of malignancy. In the present study, no sign of malignancy was detected in Warthin's tumor.