Several studies have demonstrated that the SCF-ROC1 protein is crucial for the ubiquitination of cyclin D1, D2, and D3 in humans, playing a leading role in the regulation of cyclin proteolysis [19], [24] and [32]. However, neither studies of the ROC1 immunohistochemical expression pattern nor studies comparing ROC1 and cyclin D1 expression in melanomas or other tumors are available in the literature. The expression of d-cyclins correlates with melanoma malignancy potential and prognosis. Thus, understanding the mechanism underlying d-cyclin overexpression can contribute to
the development of therapeutic approaches for melanomas overexpressing these proteins. The purpose of this work was Obeticholic Acid manufacturer to assess the relationship between ROC1 and cyclin D1 expression Nivolumab in vivo in skin melanomas and melanocytic nevi. This cross-sectional, analytic
study included 62 cases of primary skin melanoma that were allocated into four groups, according to melanoma thickness: Group 1: 15 cases of melanoma <1 mm; Group 2: 15 cases of 1.01–2 mm melanoma; Group 3: 15 cases of 2.01–4 mm melanoma; and Group 4: 17 cases of melanoma >4 mm. A total of 58 cases of compound melanocytic nevus were used as controls (Group 5). The melanoma cases did not originate from melanocytic nevi nor did they show histological regression. The sample calculus was based on the prevalence of skin melanomas in the general population. Tissue sections 4 μm thick were Oxalosuccinic acid cut, mounted on slides previously treated with poly-d-lysine, and immunostained according to the ABC technique. Incubation with primary antibodies ROC1 (clone RB-069-P, LABVISION, Westinghouse, USA; 1/800 dilution) and cyclin D1 (clone RBT14, BioSB, Santa Barbara, USA; 1/100 dilution) was carried out. The reaction was developed with DAB (Sigma
Chemical Co., St. Louis, USA) for five minutes and counterstained with Giemsa [25]. Squamous epithelium of tonsil was used as a positive control for ROC1 immunolabeling, and normal breast tissue was used as the control for cyclin D1. A semiquantitative scoring system was used for the assessment of immunohistochemical staining. Cell nuclei are either positive or negative for ROC1 and cyclin D1. The percentage of tumor cells with positive staining was determined and classified into four classes: (1) 0–25% of cells stained; (2) 26–50% of cells stained; (3) 51–75% of cells stained; and (4) 76–100% of cells stained [27].