PMID- 26657266 OWN - NLM STAT- MEDLINE DCOM- 20180326 LR - 20210217 IS - 1535-9484 (Electronic) IS - 1535-9476 (Print) IS - 1535-9476 (Linking) VI - 16 IP - 7 DP - 2017 Jul TI - Lowering Endogenous Cathepsin D Abundance Results in Reactive Oxygen Species Accumulation and Cell Senescence. PG - 1217-1232 LID - 10.1074/mcp.M115.050179 [doi] AB - Cathepsin D is reportedly to be closely associated with tumor development, migration, and invasion, but its pathological mechanism is not fully elucidated. We aimed to evaluate phenotypic changes and molecular events in response to cathepsin D knockdown. Lowering endogenous cathepsin D abundance (CR) induced senescence in HeLa cells, leading to reduced rate of cell proliferation and impaired tumorigenesis in a mouse model. Quantitative proteomics revealed that compared with control cells (EV), the abundances of several typical lysosomal proteases were decreased in the lysosomal fraction in CR cells. We further showed that cathepsin D knockdown caused increased permeability of lysosomal membrane and reactive oxygen species accumulation in CR cells, and the scavenging of reactive oxygen species by antioxidant was able to rescue cell senescence. Despite the increased reactive oxygen species, the proteomic data suggested a global reduction of redox-related proteins in CR cells. Subsequent analysis indicated that the transcriptional activity of nuclear factor erythroid-related factor 2 (Nrf2), which regulates the expression of groups of antioxidant enzymes, was down-regulated by cathepsin D knockdown. Importantly, Nrf2 overexpression significantly reduced cell senescence. Although transient oxidative stress promoted the accumulation of Nrf2 in the nucleus, we showed that the Nrf2 protein exited nucleus if oxidative stress persisted. In addition, when cathepsin D was transiently knocked down, the cathepsin-related events followed a sequential order, including lysosomal leakage during the early stage, followed by oxidative stress augmentation, and ultimately Nrf2 down-regulation and senescence. Our results suggest the roles of cathepsin D in cancer cells in maintaining lysosomal integrity, redox balance, and Nrf2 activity, thus promoting tumorigenesis. The MS Data are available via ProteomeXchange with identifier PXD002844. CI - (c) 2017 by The American Society for Biochemistry and Molecular Biology, Inc. FAU - Su, Siyuan AU - Su S AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. FAU - Zhu, Xu AU - Zhu X AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. FAU - Lin, Liang AU - Lin L AD - paragraph signProteomics Division, BGI-Shenzhen, Shenzhen, Guangdong, China, 518083. FAU - Chen, Xianwei AU - Chen X AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. FAU - Wang, Yang AU - Wang Y AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. FAU - Zi, Jin AU - Zi J AD - paragraph signProteomics Division, BGI-Shenzhen, Shenzhen, Guangdong, China, 518083. FAU - Dong, Yusheng AU - Dong Y AD - ||Beijing Protein Innovation, Beijing, China, 101318. FAU - Xie, Yingying AU - Xie Y AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. FAU - Zhu, Yinghui AU - Zhu Y AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. FAU - Zhang, Ju AU - Zhang J AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. FAU - Zhu, Jianhui AU - Zhu J AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. FAU - Xu, Dan AU - Xu D AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101. FAU - Xu, Ningzhi AU - Xu N AD - **Laboratory of Cell and Molecular Biology, Cancer Institute and Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China, 100021. FAU - Lou, Xiaomin AU - Lou X AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101; siqiliu@big.ac.cn louxm@big.ac.cn. FAU - Liu, Siqi AU - Liu S AD - From the double daggerCAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China, 100101; siqiliu@big.ac.cn louxm@big.ac.cn. AD - section signUniversity of Chinese Academy of Sciences, Beijing, China, 100049. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151210 PL - United States TA - Mol Cell Proteomics JT - Molecular & cellular proteomics : MCP JID - 101125647 RN - 0 (NF-E2-Related Factor 2) RN - 0 (NFE2L2 protein, human) RN - 0 (Reactive Oxygen Species) RN - EC 3.4.23.5 (CTSD protein, human) RN - EC 3.4.23.5 (Cathepsin D) SB - IM MH - A549 Cells MH - Animals MH - Cathepsin D/*genetics/metabolism MH - Cell Nucleus/genetics/metabolism MH - Cell Proliferation MH - Cellular Senescence MH - Down-Regulation MH - Female MH - Gene Knockdown Techniques MH - HeLa Cells MH - Humans MH - Lysosomes/genetics/metabolism MH - Mice MH - NF-E2-Related Factor 2/*metabolism MH - Neoplasm Transplantation MH - Neoplasms/genetics/*metabolism MH - Oxidative Stress MH - Proteomics/*methods MH - Reactive Oxygen Species/*metabolism PMC - PMC5500756 EDAT- 2015/12/15 06:00 MHDA- 2018/03/27 06:00 PMCR- 2018/07/01 CRDT- 2015/12/15 06:00 PHST- 2015/04/16 00:00 [received] PHST- 2015/11/25 00:00 [revised] PHST- 2015/12/15 06:00 [pubmed] PHST- 2018/03/27 06:00 [medline] PHST- 2015/12/15 06:00 [entrez] PHST- 2018/07/01 00:00 [pmc-release] AID - S1535-9476(20)34185-2 [pii] AID - M115.050179 [pii] AID - 10.1074/mcp.M115.050179 [doi] PST - ppublish SO - Mol Cell Proteomics. 2017 Jul;16(7):1217-1232. doi: 10.1074/mcp.M115.050179. Epub 2015 Dec 10.