1. Academic Validation
  2. S1P/S1PR3 axis promotes aerobic glycolysis by YAP/c-MYC/PGAM1 axis in osteosarcoma

S1P/S1PR3 axis promotes aerobic glycolysis by YAP/c-MYC/PGAM1 axis in osteosarcoma

  • EBioMedicine. 2019 Feb;40:210-223. doi: 10.1016/j.ebiom.2018.12.038.
Yifei Shen 1 Shujie Zhao 2 Shenyu Wang 3 Xiaohui Pan 1 Yunkun Zhang 1 Jingwen Xu 4 Yuqing Jiang 1 Haibo Li 1 Qiang Zhang 1 Jianbo Gao 4 Qin Yang 5 Yang Zhou 6 Shuheng Jiang 5 Huilin Yang 3 Zhigang Zhang 5 Rong Zhang 7 Jun Li 8 Dong Zhou 9
Affiliations

Affiliations

  • 1 Department of Orthopedics, The Affiliated Hospital of Nanjing Medical University, Changzhou No. 2 People's Hospital, Changzhou, Jiangsu 213003, China.
  • 2 Department of Orthopedic, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210000, China.
  • 3 Department of Orthopedics, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, China.
  • 4 Department of Nutrition, The Affiliated Hospital of Nanjing Medical University, Changzhou No. 2 People's Hospital, Changzhou, Jiangsu 213003, China.
  • 5 State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200240, China.
  • 6 Department of General Surgery, the First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu 210029, China.
  • 7 Department of Obstetrics and Gynecology, Fengxian Hospital, Southern Medical University, Shanghai 201499, China.. Electronic address: Rongzhang@163.com.
  • 8 State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: Junli@shsci.org.
  • 9 Department of Orthopedics, The Affiliated Hospital of Nanjing Medical University, Changzhou No. 2 People's Hospital, Changzhou, Jiangsu 213003, China. Electronic address: zhoudong1012@163.com.
Abstract

Background: Osteosarcoma (OS) is a malignant tumor mainly occurring in young people. Due to the limited effective therapeutic strategies, OS patients cannot achieve further survival improvement. G-protein-coupled receptors (GPCRs) constitute the largest family of cell membrane receptors and consequently hold the significant promise for tumor imaging and targeted therapy. We aimed to explore the biological functions of Sphingosine 1-phosphate receptor 3 (S1PR3), one of the members of GPCRs family, in OS and the possibility of S1PR3 as an effective target for the treatment of osteosarcoma.

Methods: The quantitative real time PCR (qRT-PCR) and western blotting were used to analyze the mRNA and protein expressions. Cell counting kit-8 (CCK8), colony formation assay and cell Apoptosis assay were performed to test the cellular proliferation in vitro. Subcutaneous xenograft mouse model was generated to evaluate the functions of S1PR3 in vivo. RNA Sequencing was used to compare gene expression patterns between S1PR3-knockdown and control MNNG-HOS cells. In addition, metabolic alternations in OS cells were monitored by XF96 metabolic flux analyzer. Co-immunoprecipitation (Co-IP) assay was used to explore the interaction between Yes-associated protein (YAP) and c-Myc. Chromatin immunoprecipitation was used to investigate the binding capability of PGAM1 and YAP or c-Myc. Moreover, the activities of promoter were determined by the luciferase reporter assay.

Findings: S1PR3 and its specific ligand Sphingosine 1-phosphate (S1P) were found elevated in OS, and the higher expression of S1PR3 was correlated with the poor survival rate. Moreover, our study has proved that the S1P/S1PR3 axis play roles in proliferation promotion, Apoptosis inhibition, and aerobic glycolysis promotion of osteosarcoma cells. Mechanistically, the S1P/S1PR3 axis inhibited the phosphorylation of YAP and promoted the nuclear translocation of YAP, which contributed to the formation of the YAP-c-MYC complex and enhanced transcription of the important glycolysis Enzyme PGAM1. Moreover, the S1PR3 Antagonist TY52156 exhibited in vitro and in vivo synergistic inhibitory effects with methotrexate on OS cell growth.

Interpretation: Our study unveiled a role of S1P, a bioactive phospholipid, in glucose metabolism reprogram through interaction with its receptor S1PR3. Targeting S1P/S1PR3 axis might serve as a potential therapeutic target for patients with OS. FUND: This research was supported by National Natural Science Foundation of China (81472445 and 81672587).

Keywords

Cell proliferation; Osteosarcoma; S1P; S1PR3; Warburg effect; YAP.

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