Immunohistological comparison of the World Health Organization (WHO) and Ljubljana classifications on the grading of preneoplastic lesions of the larynx

Abstract

There is still controversy about the classification of laryngeal preneoplastic lesions. In this study, we compared the World Health Organization (WHO) and Ljubljana histological classifications with regard to laryngeal epithelial hyperplastic-dysplastic lesions in a single series to determine their relation to statistically significant criteria when the mechanisms of neoplastic progression are considered. Emphasis was put on p53 expression, proliferative activity, and angiogenesis. Fifty-four laryngeal biopsies with preneoplastic changes were re-evaluated and classified according to both classifications. The streptavidin-biotin method was used for immunohistochemical staining for Ki-67, p53, and CD34 antibodies. A positive correlation was obtained between the histological categories and Ki-67, p53, and CD34 expressions using both classifications (Spearman's Correlation test). There was a significant difference between the histological categories of both the WHO and the Ljubljana classifications, when the expression of the three markers was compared (Kruskal Wallis test, p = 0.000 for each). Further evaluation revealed a statistically significant difference between all categories of both classifications, excluding the p53 overexpression scores and vessel counts in mild and moderate dysplasia categories (Mann-Whitney U-test, p = 0.209, and 0.091 respectively), and the p53 overexpression scores in severe dysplasia and carcinoma in situ categories (Mann-Whitney U-test, p = 0.249) of the WHO classification. Similarly, no significant differences were found between severe dysplasia and carcinoma in situ, as well as between atypical hyperplasia and carcinoma in situ categories using both classifications for the Ki-67 expression (Mann-Whitney U-test, p = 0.806, and 0.111, respectively). Our results suggest that regarding the mechanisms of neoplastic progression such as p53 expression and angiogenesis, the histological categories of the Ljubljana classification seem to depend on additional evidence. Therefore, we support the use of the Ljubljana classification.