To evaluate the roles of α9-nAChR in the proliferation, migration and PD-L1 upregulation of melanoma cells, we generated stable α9-nAChR-siRNA-expressing A2058 cells, α9-nAChR-overexpressing A2058 cells and α9-nAChR-overexpressing MDA-MB 435 cells by transfection with α9-nAChR-small interfering RNA (siRNA) or pcDNA3.1-α9-nAChR overexpression vectors. Here, CD274 is linked to melanoma.