Objective: To study, under controlled conditions, the applicability of automated image analysis of immunohistochemical markers as an indicator of development and progression in tobacco component-induced tumors in the respiratory tract.
Study design: Amount, location, size, shape and intensity of staining of proliferating cell and p53 antigen in chemically induced precursors and squamous cell carcinoma of the hamster lung were determined by computer-assisted morphometry.
Results: The total expression of proliferating cell nuclear antigen (PCNA) and p53 expression increased consistently during the formation of papillomas and squamous cell carcinomas of the larynx, trachea, bronchi and lungs. Individual preneoplastic cells in epithelial dysplasia expressed PCNA staining, increasing with increasing cell size and optical density, indicating antibody- staining intensity, in relation to the increased degree of cellular atypia. In malignant tumors, cell size decreased with decreasing differentiation, while antibody staining intensity remained unchanged. The increased alterations in cell shape and percent PCNA-positive cells observed in dysplastic epithelium and squamous cell carcinomas were statistically significant using Spearman's correlation coefficient. Squamous cell carcinomas consisted of two tumor cell populations with different cell shapes, and PCNA and p53 staining intensity. Altering measurement conditions-antibody threshold levels, size of measured area and repeating measurements-showed computer-assisted image analysis to give sensitive, reliable and consistent results.
Conclusion: Computer-assisted analysis of immunohistochemical staining showed high sensitivity and reproducibility; however, the results depended upon the method of study.