autophagy impairment induces premature senescence in primary human fibroblasts人类成纤维细胞自噬缺陷导致过早衰老在初级.pdfVIP

Autophagy Impairment Induces Premature Senescence in Primary Human Fibroblasts 2 1 1 1 1,2,3 Hyun Tae Kang , Ki Baek Lee , Sung Young Kim , Hae Ri Choi , Sang Chul Park * 1 Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, Korea, 2 Institute on Aging, Seoul National University College of Medicine, Seoul, Korea, 3 Lee Gil Ya Cancer and Diabetes Institute, Gachon University of Medicine and Science, Incheon, Korea Abstract Background: Recent studies have demonstrated that activation of autophagy increases the lifespan of organisms from yeast to flies. In contrast to the lifespan extension effect in lower organisms, it has been reported that overexpression of unc-51- like kinase 3 (ULK3), the mammalian homolog of autophagy-specific gene 1 (ATG1), induces premature senescence in human fibroblasts. Therefore, we assessed whether the activation of autophagy would genuinely induce premature senescence in human cells. Methodology/Principal Findings: Depletion of ATG7, ATG12, or lysosomal-associated membrane protein 2 (Lamp2) by transfecting siRNA or infecting cells with a virus containing gene-specific shRNA resulted in a senescence-like state in two strains of primary human fibroblasts. Prematurely senescent cells induced by autophagy impairment exhibited the senescent phenotypes, similar to the replicatively senescent cells, such as increased senescence associated b-galactosidase (SA-b-gal) activity, reactive oxygen species (ROS) generation, and accumulation of lipofuscin. In addition, expression levels of ribosomal protein S6 kinase1 (S6K1), p-S6K1, p-S6, and