Background and aim Benign hepatic nodular lesions mimicking hepatocellular carcinoma (HCC) may be categorized into hyperplastic nodular lesions, neoplastic nodular lesions, and miscellaneous nodular lesions. This study aimed to assess the role of glypican 3 (GPC3), arginase 1 (ARG-1), and CD34 in differentiation between benign and malignant primary hepatic lesions.Materials and methods This study included 48 cases, where 25 (52.08%) cases were diagnosed as primary HCC in the liver and 23 (47.92%) cases were diagnosed as benign hepatic lesions. Of the 48 patients, five (10.42%) cases were diagnosed as focal nodular hyperplasia, four (8.33%) cases were diagnosed as hepatic adenoma, eight (16.67%) cases were diagnosed as dysplastic nodule, and six (12.50%) cases were diagnosed as regenerative nodule.Results The sensitivity of GPC3 for differentiation between benign and malignant primary hepatic lesions was 80% and its specificity was 82.5%. The sensitivity of ARG-1 was 90%, and its specificity was 0.0%. The sensitivity of CD34 for HCC in the study group was 84% and its specificity was 91.3%.Conclusion There was high sensitivity and specificity of both GPC3 and CD34 immunostaining for distinction of primary HCC from benign hepatic mimicker lesions, whereas there was high sensitivity of ARG-1 in both benign and malignant primary hepatic lesions but with no specificity for differentiating the benign from malignant primary hepatic lesions.