A p16-Ki-67-HMB45 immunohistochemistry scoring system as an ancillary diagnostic tool in the diagnosis of melanoma

Arnaud Uguen, Matthieu Talagas, Sebastian Costa, Sandrine Duigou, Stéphanie Bouvier, Marc De Braekeleer, Pascale Marcorelles

Melanoma is a skin cancer which treatment requires early diagnosis and large surgical removal. The histopathological diagnosis of a melanocytic tumour is sometimes difficult between a benign nevus and a malignant melanoma. We built an immunomarker-based score to differentiate nevi from melanomas. Two independent sets of 308 (first set) and 62 (validation set) formalin-fixed and paraffin embedded tumour samples were studied using p16-Ki-67 and HMB45-MelanA dual-staining immunohistochemistry. In the first set of tumours, high Ki-67 index, low to null p16 immunohistochemistry and absence of HMB45 immunohistochemistry gradient were more frequent in melanomas (156 primary tumours and 78 metastases) than in nevi (74 tumours). Nevertheless, none of these single parameters was able to differentiate all primary melanomas from all nevi. We built a scoring system based on the addition of semi-quantitative scorings of Ki-67 (0: <2 %; 1:2-5 %; 2:6-10 %, 3:11-20 %; 4:>20 %) and p16 (0:>50 % stained cells; 1:11-50 %; 2:1-10 %; 3:0 %) and HMB45 staining (0: gradient present; 1: doubtful/inconclusive gradient; 2: gradient absent). A p16-Ki-67-HMB45 total score from 0 to 9 permitted to classify nevi (score <4) and primary melanomas (score ≥4) with a sensitivity of 97.4 % and a specificity of 97.3 % in the first set of tumours. Sensibility and specificity of 100 % were obtained in a second set (validation set) of 62 tumours (46 melanomas and 16 nevi). The total scoring also allowed analyzing 11 difficult or initially misdiagnosed tumours in our files. We propose a valuable triple p16-Ki-67-HMB45 immunohistochemistry scoring system to help pathologists in the differential diagnosis of melanomas and nevi.