PMID- 32859246 OWN - NLM STAT- MEDLINE DCOM- 20210607 LR - 20210607 IS - 1756-9966 (Electronic) IS - 0392-9078 (Print) IS - 0392-9078 (Linking) VI - 39 IP - 1 DP - 2020 Aug 28 TI - Lysine-222 succinylation reduces lysosomal degradation of lactate dehydrogenase a and is increased in gastric cancer. PG - 172 LID - 10.1186/s13046-020-01681-0 [doi] LID - 172 AB - BACKGROUND: Lysine succinylation is an emerging posttranslational modification that has garnered increased attention recently, but its role in gastric cancer (GC) remains underexplored. METHODS: Proteomic quantification of lysine succinylation was performed in human GC tissues and adjacent normal tissues by mass spectrometry. The mRNA and protein levels of lactate dehydrogenase A (LDHA) in GC and adjacent normal tissues were analyzed by qRT-PCR and western blot, respectively. The expression of K222-succinylated LDHA was measured in GC tissue microarray by the K222 succinylation-specific antibody. The interaction between LDHA and sequestosome 1 (SQSTM1) was measured by co-immunoprecipitation (co-IP) and proximity ligation assay (PLA). The binding of carnitine palmitoyltransferase 1A (CPT1A) to LDHA was determined by co-IP. The effect of K222-succinylated LDHA on tumor growth and metastasis was evaluated by in vitro and in vivo experiments. RESULTS: Altogether, 503 lysine succinylation sites in 303 proteins were identified. Lactate dehydrogenase A (LDHA), the key enzyme in Warburg effect, was found highly succinylated at K222 in GC. Intriguingly, this modification did not affect LDHA ubiquitination, but reduced the binding of ubiquitinated LDHA to SQSTM1, thereby decreasing its lysosomal degradation. We demonstrated that CPT1A functions as a lysine succinyltransferase that interacts with and succinylates LDHA. Moreover, high K222-succinylation of LDHA was associated with poor prognosis in patients with GC. Finally, overexpression of a succinylation-mimic mutant of LDHA promoted cell proliferation, invasion, and migration. CONCLUSIONS: Our data revealed a novel lysosomal pathway of LDHA degradation, which is mediated by the binding of K63-ubiquitinated LDHA to SQSTM1. Strikingly, CPT1A succinylates LDHA on K222, which thereby reduces the binding and inhibits the degradation of LDHA, as well as promotes GC invasion and proliferation. This study thus uncovers a new role of lysine succinylation and the mechanism underlying LDHA upregulation in GC. FAU - Li, Xiang AU - Li X AD - Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, 510095, China. AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. AD - Nanjing University of Chinese Medicine, Nanjing, 210023, China. FAU - Zhang, Chen AU - Zhang C AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Zhao, Ting AU - Zhao T AD - Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, 510095, China. FAU - Su, Zhongping AU - Su Z AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Li, Mengjing AU - Li M AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Hu, Jiancheng AU - Hu J AD - Division of Cellular and Molecular Research, National Cancer Centre Singapore, Singapore, 169610, Singapore. AD - Cancer and Stem Cell Biology Program, Duke-NUS Medical School, Singapore, 169857, Singapore. FAU - Wen, Jianfei AU - Wen J AD - The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China. FAU - Shen, Jiajia AU - Shen J AD - The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China. FAU - Wang, Chao AU - Wang C AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Pan, Jinshun AU - Pan J AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Mu, Xianmin AU - Mu X AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Ling, Tao AU - Ling T AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Li, Yingchang AU - Li Y AD - Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, 510095, China. FAU - Wen, Hao AU - Wen H AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. FAU - Zhang, Xiaoren AU - Zhang X AD - Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, 510095, China. AD - Key Laboratory of Cell Homeostasis and Cancer Research of Guangdong Higher Education Institutes, Guangzhou Medical University, Guangzhou, 510182, China. FAU - You, Qiang AU - You Q AUID- ORCID: 0000-0003-3308-4127 AD - Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, 510095, China. qiang.you@live.com. AD - Department of Biotherapy, Department of Surgery, Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, China. qiang.you@live.com. AD - Key Laboratory of Cell Homeostasis and Cancer Research of Guangdong Higher Education Institutes, Guangzhou Medical University, Guangzhou, 510182, China. qiang.you@live.com. LA - eng GR - 81671543/National Natural Science Foundation of China/ GR - 81870409/National Natural Science Foundation of China/ GR - 81802464/National Natural Science Foundation of China/ PT - Journal Article DEP - 20200828 PL - England TA - J Exp Clin Cancer Res JT - Journal of experimental & clinical cancer research : CR JID - 8308647 RN - 0 (Biomarkers, Tumor) RN - 0 (SQSTM1 protein, human) RN - 0 (Sequestosome-1 Protein) RN - AB6MNQ6J6L (Succinic Acid) RN - EC 1.1.1.27 (L-Lactate Dehydrogenase) RN - EC 1.1.1.27 (LDHA protein, human) RN - EC 2.3.1.21 (CPT1A protein, human) RN - EC 2.3.1.21 (Carnitine O-Palmitoyltransferase) RN - K3Z4F929H6 (Lysine) SB - IM MH - Animals MH - Apoptosis MH - Biomarkers, Tumor/genetics/*metabolism MH - Carnitine O-Palmitoyltransferase/genetics/metabolism MH - Cell Proliferation MH - Female MH - Gene Expression Regulation, Enzymologic MH - Gene Expression Regulation, Neoplastic MH - Humans MH - L-Lactate Dehydrogenase/chemistry/genetics/*metabolism MH - Lung Neoplasms/genetics/metabolism/secondary MH - Lysine/*chemistry MH - Lysosomes/*metabolism MH - Male MH - Melanoma/genetics/metabolism/pathology MH - Mice MH - Mice, Nude MH - Middle Aged MH - Prognosis MH - *Protein Processing, Post-Translational MH - Proteolysis MH - Sequestosome-1 Protein/genetics/metabolism MH - Stomach Neoplasms/genetics/metabolism/*pathology MH - Succinic Acid/*chemistry MH - Survival Rate MH - Tumor Cells, Cultured MH - Xenograft Model Antitumor Assays PMC - PMC7455916 OTO - NOTNLM OT - CPT1A OT - Gastric cancer OT - LDHA OT - Lysine succinylation OT - SQSTM1 COIS- The authors declare that they have no competing interests. EDAT- 2020/08/30 06:00 MHDA- 2021/06/08 06:00 PMCR- 2020/08/28 CRDT- 2020/08/30 06:00 PHST- 2020/05/17 00:00 [received] PHST- 2020/08/17 00:00 [accepted] PHST- 2020/08/30 06:00 [entrez] PHST- 2020/08/30 06:00 [pubmed] PHST- 2021/06/08 06:00 [medline] PHST- 2020/08/28 00:00 [pmc-release] AID - 10.1186/s13046-020-01681-0 [pii] AID - 1681 [pii] AID - 10.1186/s13046-020-01681-0 [doi] PST - epublish SO - J Exp Clin Cancer Res. 2020 Aug 28;39(1):172. doi: 10.1186/s13046-020-01681-0.