earticle

영어

Cell death which is very important in mammals, is orchestrated by two types of death signaling: the Apoptosis (Programmed cell death) and Necroptosis (Programmed necrosis). Apoptosis and Necroptosis share common proteins such as FADD, Caspase-8 and RIP1 kinase in their molecular mechanism. However, only Necroptosis includes RIP3 Kinase dependent signaling. Therefore, RIP3 Kinase is a key determinator of necroptotic cell death. RIP3 kinase which is most important protein in necroptosis is strongly down-regulated their expression in most cancer cell. It's known that DNMT1 (DNA methyltransferase 1) plays an important roles in this process. RIP3 promoter is methylated by DNMT1 in most cancer cell. On the other hand, O-GlcNAc modification is a single sugar modification occurs in cytosol and nucleus. This modification has many roles in cellular responses including nutrient sensing. Here, we identified that cellular global O-GlcNAcylation change affects to RIP3 kinase expression by real time RT-PCR. In cancer HeLa and HepG2, mRNA level of RIP3 kinase is largely increase when OGA inhibitor Thiamet-G was treated. We also confirm that OGA inhibitor Thiamet-G induced the increase of endogenouse RIP3 kinase protein level in HEK293 and HeLa cell by Western blotting. And we will confirm the O-GlcNAcylation of RIP3 kinase using immunoprecipitation and succinylated wheat germ agglutinin (sWGA) precipitation. Ultimately we aim to find the physiological functions of cellular O-GlcNAcylation change in RIP3-mediated necroptosis.