ORIGINAL PAPER Analysis of Immunohistochemical Expression of K19 in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma Using Color Deconvolution-Image Analysis Method

Abstract K19 is an intermediate filament protein that has been investigated in oral squamous cell carcinoma (OSCC), but that has not been correlated with the amount of keratin produced within well-differentiated OSCC grade. The aim of the present study was to objectively analyze K19 immuno-expression in OSCC and to validate the utility of K19 in dif-ferentiation among grades of oral epithelial dysplasia (OED). Formalin-fixed tissues of 36 primary OSCC (22 well, 10 moderately, 4 poorly differentiated), 43 OED (23 mild, 8 moderate, 12 severe), and 11 normal oral epithelium (NOE) were included. K19 was immunostained using HRP-DAB method. The percentage of K19-positive area was found using color deconvolution program in ImageJ image analysis system (public domain software, National Institutes of Health, Bethesda, MD, USA) and analyzed using independent sam-ples t tests and ANOVA test. K19 scores in NOE, mild, moderate and severe OED were: 1.8, 3.4, 21, and 50.3%, respectively, with significant association with the grade (t test P \ 0.05). Well-differentiated OSCC with \30 % keratin pearl formation expressed significantly higher K19 scores compared to well-differentiated OSCC with [30 % keratin pearls (28.6 and 1.2%, respectively, P \ 0.05). K19 scores in moderately and poorly differentiated OSCC were 60.8 and 61.3%, respectively. K19 scores significantly differentiated between two subgroups of tumors within well-differentiated OSCC grade and reflected histologic differentiation as well as probably predicting the clinical outcome. Combining K19 immunostain with the regular H&E stain may be helpful to facilitate and assure assigning a more accurate grade for OED