Objective: The cervical cancer is one of the most common cancers among women worldwide. Diagnosis of CIN affected by high rates of discordance among pathologists. Therefore, we need to other adjunct methods for accurate diagnosis of CIN versus benign lesions in equivocal cases. The aim of this study was evaluation of Ki-67 (MIB-1), CK17 and P16 INK 4a (P16) markers by immunohistochemical method in differentiating CIN from benign cervical lesions. Method: Seventy-seven cervical biopsies that originally diagnosed as non-CIN (n = 31) and CIN (n = 46), were reviewed by three pathologists and re-classified as non-CIN (n = 54) and CIN (n = 23), based on agreement between at least two of three, to obtain a consensus diagnosis. Consensus diagnosis was defined as the "Gold Standard". Then immunostaining for Ki67, P16 and CK17 was performed on all cases and their results were compared with original and consensus diagnosis. Results: The overall agreement between original and consensus diagnosis was 67.5 % (Kappa = 0.39, P-value < 0.001). The sensitivity and specificity of Ki67 immunostaining were 95.6 % and 85.1 % and for P16 were 91.3 and 98.1 %, respectively. The sensitivity and specificity of CK17 negative staining for CIN detection were 39.1 % and 40.7 % respectively. Conclusion: We recommended using Ki67 and P16 markers as complementary tests for differentiation between dysplastic and non-dysplastic lesions.