Fig. 5
LINC01116 Promotes Nascent Protein Synthesis and Influences Melanoma Cell Stemness. (A) Fluorescence intensity analysis demonstrated successful transfection efficiency across experimental groups. (B) CCK-8 proliferation assay revealed distinct growth dynamics: h-NC group exhibited the highest proliferation rate. sh-LINC01116 and sh-LINC01116 + Inh-NC groups showed the suppression of proliferation. (C) Clonogenic assay results demonstrated: sh-NC group displayed superior clonogenic potential. sh-LINC01116 + miR-432-5p inhibitor group exhibited partial restoration of clonogenicity compared to sh-LINC01116 and sh-LINC01116 + Inh-NC groups. (D) Cell cycle analysis via flow cytometry identified: G1/S phase arrest in sh-LINC01116-transfected cells, miR-432-5p inhibitor rescued this arrest. (E) 3D tumor spheroid assay:​sh-NC and sh-LINC01116 + miR-432-5p inhibitor groups formed larger, more compact spheroids. (F) WB analysis was performed to detect the protein levels of stemness genes Nestin, ABCB5, CD133, and ALDH1 in the A375 and sk-mel-28 cell lines across the following groups: sh-NC, sh-LINC01116, sh-LINC01116 + Inh-NC, and sh-LINC01116 + miR-432-5p inhibitor. The results showed that, with the exception of ABCB5, the expression levels of the other genes were significantly lower in the sh-LINC01116 and sh-LINC01116 + Inh-NC groups compared to the sh-NC and sh-LINC01116 + miR-432-5p inhibitor groups.
