For this purpose, stably transfected t-P2Y1, h-P2Y2, h-P2Y4, and r-P2Y6-1321N1 astrocytoma cell line was used and fluorescent based Ca2+ influx was assessed by using a Ca2+ binding dye i.e., Fura-2 a.m. The cytotoxicity component was removed using a cell viability assay (MTT assay). Here, P2RY4 is linked to astrocytoma (excluding glioblastoma).