ZnCl2 와 Calmodulin 길항제에 의한 Human Retinoblastoma Cell 의 세포사 기전에 관한 연구

In order to know the mechanismof retinoblastoma cell death by Zinc(Zn) and calmodulin antagonist, Y79 retinoblastoma was treated with ZnCl2 and calmodulin antagonist(W13). Using electron microscopic study and biochemical assay, the Zn-induced and W13-induced cell death were analyzed. In addition, western blotting and Raspull down assay were done to detect the signal changes during retinoblastoma cell death. ZnCl2 treatment caused extensive cell death within 16 hours. Morphological and biochemical analysis revealed that Zn-induced cell death was necrosis. It seemed that mitogen activated protein(MAP) kinase or phophoinositide-3-kinase(PI3K) was not involved in Zn-induced retinoblastoma cell death. However, protein kinase C potenticated Zn-induced cell death. Calmodulin antagonist, W13, dramatically induced retinoblastoma cell death after 16 hours of treatment. Apoptotic processes, such as p38 MAP kinase was activated after W13 treatment, and the cell death was partially inhibited by the cotreatment of p38 kinase inhibitor. Rac, and upstream molecule of p38 kianse, activity was increased with the treatment of W13. These findings indicate that oxydative stress induces necrosis but W13 treatment results in apoptosis in retinoblastoma cell. In addition, it seems that Rac/p38 kinase may be involved in W13-induced apoptosis of retinoblastoma cells(J Korean Ophthalmol Soc 41:2686~2698, 2000).