S1P/S1PR3 axis promotes aerobic glycolysis by YAP/c-MYC/PGAM1 axis in osteosarcoma.
Adaptor Proteins, Signal Transducing
/ metabolism
Animals
Apoptosis
/ genetics
Bone Neoplasms
/ genetics
Cell Line, Tumor
Disease Models, Animal
Gene Expression Regulation, Neoplastic
Gene Knockdown Techniques
Glycolysis
Heterografts
Humans
Immunohistochemistry
Lysophospholipids
/ metabolism
Male
Mice
Multiprotein Complexes
/ metabolism
Osteosarcoma
/ genetics
Oxidative Phosphorylation
Phosphoglycerate Mutase
/ metabolism
Phosphoproteins
/ metabolism
Protein Binding
Proto-Oncogene Proteins c-myc
/ metabolism
Receptors, Lysosphingolipid
/ antagonists & inhibitors
Signal Transduction
Sphingosine
/ analogs & derivatives
Sphingosine-1-Phosphate Receptors
Transcription Factors
YAP-Signaling Proteins
Cell proliferation
Osteosarcoma
S1P
S1PR3
Warburg effect
YAP
Journal
EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039
Informations de publication
Date de publication:
Feb 2019
Feb 2019
Historique:
received:
07
11
2018
revised:
18
12
2018
accepted:
18
12
2018
pubmed:
28
12
2018
medline:
27
6
2019
entrez:
28
12
2018
Statut:
ppublish
Résumé
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. 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. 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. 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).
Sections du résumé
BACKGROUND
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
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
RESULTS
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
CONCLUSIONS
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).
Identifiants
pubmed: 30587459
pii: S2352-3964(18)30615-7
doi: 10.1016/j.ebiom.2018.12.038
pmc: PMC6412077
pii:
doi:
Substances chimiques
Adaptor Proteins, Signal Transducing
0
Lysophospholipids
0
Multiprotein Complexes
0
Phosphoproteins
0
Proto-Oncogene Proteins c-myc
0
Receptors, Lysosphingolipid
0
Sphingosine-1-Phosphate Receptors
0
Transcription Factors
0
YAP-Signaling Proteins
0
YAP1 protein, human
0
sphingosine-1-phosphate receptor-3, human
0
sphingosine 1-phosphate
26993-30-6
Phosphoglycerate Mutase
EC 5.4.2.11
phosphoglycerate mutase 1, human
EC 5.4.2.11
Sphingosine
NGZ37HRE42
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
210-223Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.
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