sirt1 overexpression antagonizes cellular senescence with activated erks6k1 signaling in human diploid fibroblastssirt1过度对抗细胞衰老与人类二倍体成纤维细胞激活erks6k1信号.pdfVIP

SIRT1 Overexpression Antagonizes Cellular Senescence with Activated ERK/S6k1 Signaling in Human Diploid Fibroblasts Jing Huang1,2, Qini Gan1,2, Limin Han1,2, Jian Li1,2, Hai Zhang1,2, Ying Sun1,2, Zongyu Zhang1,2, Tanjun Tong1,2* 1 Peking University Research Center on Aging, Beijing, People’s Republic of China, 2 Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing, People’s Republic of China Abstract Sir2, a NAD-dependent deacetylase, modulates lifespan in yeasts, worms and flies. The SIRT1, mammalian homologue of Sir2, regulates signaling for favoring survival in stress. But whether SIRT1 has the function to influence cell viability and senescence under non-stressed conditions in human diploid fibroblasts is far from unknown. Our data showed that enforced SIRT1 expression promoted cell proliferation and antagonized cellular senescence with the characteristic features of delayed Senescence-Associated b-galactosidase (SA-b-gal) staining, reduced Senescence-Associated Heterochromatic Foci (SAHF) formation and G1 phase arrest, increased cell growth rate and extended cellular lifespan in human fibroblasts, while dominant- negative SIRT1 allele (H363Y) did not significantly affect cell growth and senescence but displayed a bit decreased lifespan.. Western blot results showed that SIRT1 reduced the expression of p16INK4A and promoted phosphorylation of Rb. Our data also exposed that overexpression of SIRT1 was accompanied by enhanced activation of ERK and S6K1 signaling. These effects were mimicked in both WI38 cells and 2BS cells by concentration-dependent resveratrol, a SIRT1 activator. It was noted that treatment of SIRT1-.transfected cells with Rapamycin, a mTOR inhibitor, reduced the phosphorylation of S6K1 and the expressio