circTLK1 sequence containing miR-495-3p wild-type or mutant binding site was cloned into pmirGLO vector (LMAI Bio, Shanghai, China) to form WT-circTLK1 and MUT-circTLK1 vectors. Meanwhile, CBL 3′UTR harboring miR-495-3p wild-type or mutant binding site was inserted into pmirGLO vector (LMAI Bio) to form WT-CBL 3′UTR and MUT-CBL 3′UTR vectors. Next, the constructed vector and miR-NC mimic or miR-495-3p mimic was co-transfected into Caki-1 and 786-O cells. Subsequently, the luciferase intensity was detected via Dual-Lucy Assay Kit (Solarbio).

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