Retinoblastoma-Binding Protein 5 Regulates H3K4 Methylation Modification to Inhibit the Proliferation of Melanoma Cells by Inactivating the Wnt/β-Catenin and Epithelial-Mesenchymal Transition Pathways

Figure 2

RBBP5 was downregulated in melanoma. (a, b) Western blot to explore the expression of RBBP5 in three pairs of melanoma tissue compared with nevi and adjacent tissue. (c, d) Western blotting and (e) qRT-PCR were performed to verify the mRNA and protein expression of RBBP5 in two melanoma cell lines (A375 and A2058) compared with a human immortalized keratinocyte (HaCaT) cell line. (f) IHC of RBBP5 was performed in 106 cases of melanoma tumor tissues, 100 cases of adjacent tissues, and 23 cases of nevi tissues. Representative images with different tissues stained are shown. Scale bar: 200 µm and 50 µm. (g, h) The relative score of RBBP5 was quantified by the grade of staining intensity and the percentage of positively stained cells. A375 and A2058 cells were transfected with empty vector (shRBBP5-vector) or shRNAs targeting RBBP5 (shRBBP5), and the relative mRNA level of RBBP5 was measured by qRT-PCR. GAPDH was used as an internal control (j). The protein expression of RBBP5 was detected by western blotting. GAPDH was used as a loading control (k, l). A375 and A2058 cells were transfected with RBBP5 empty vector (RBBP5-vector) or RBBP5 overexpression lentivirus (RBBP5), and transfection efficiency was verified by q-PCR (i) and western blotting (m, n). GAPDH was used as a loading control. Data are represented as mean ± SD of three independent experiments. ; ns, no significance.