Loss of RACK1 promotes glutamine addiction via activating AKT/mTOR/ASCT2 axis to facilitate tumor growth in gastric cancer.

ASCT2 Autophagy Gastric cancer Glutamine addiction RACK1

Journal

Cellular oncology (Dordrecht)
ISSN: 2211-3436
Titre abrégé: Cell Oncol (Dordr)
Pays: Netherlands
ID NLM: 101552938

Informations de publication

Date de publication:
14 Aug 2023
Historique:
accepted: 31 07 2023
medline: 14 8 2023
pubmed: 14 8 2023
entrez: 14 8 2023
Statut: aheadofprint

Résumé

Metabolic reprogramming is closely related to the development of gastric cancer (GC), which remains as the fourth leading cause of cancer-related death worldwide. As a tumor suppressor for GC, whether receptor for activated C-kinase 1 (RACK1) play a modulatory role in metabolic reprogramming remains largely unclear. GC cell lines and cell-derived xenograft mouse model were used to identify the biological function of RACK1. Flow cytometry and Seahorse assays were applied to examine cell cycle and oxygen consumption rate (OCR), respectively. Western blot, real-time PCR and autophagy double fluorescent assays were utilized to explore the signaling. Immunohistochemistry was performed to detect the expression of RACK1 and other indicators in tissue sections. Loss of RACK1 facilitated the viability, colony formation, cell cycle progression and OCR of GC cells in a glutamine-dependent manner. Further investigation revealed that RACK1 knockdown inhibited the lysosomal degradation of Alanine-serine-cysteine amino acid transporter 2 (ASCT2). Mechanistically, depletion of RACK1 remarkably decreased PTEN expression through up-regulating miR-146b-5p, leading to the activation of AKT/mTOR signaling pathway which dampened autophagy flux subsequently. Moreover, knockdown of ASCT2 could reverse the promotive effect of RACK1 depletion on GC tumor growth both in vitro and in vivo. Tissue microarray confirmed that RACK1 was negatively correlated with the expression of ASCT2 and p62, as well as the phosphorylation of mTOR. Together, our results demonstrate that the suppressive function of RACK1 in GC is associated with ASCT2-mediated glutamine metabolism, and imply that targeting RACK1/ASCT2 axis provides potential strategies for GC treatment.

Sections du résumé

BACKGROUND BACKGROUND
Metabolic reprogramming is closely related to the development of gastric cancer (GC), which remains as the fourth leading cause of cancer-related death worldwide. As a tumor suppressor for GC, whether receptor for activated C-kinase 1 (RACK1) play a modulatory role in metabolic reprogramming remains largely unclear.
METHODS METHODS
GC cell lines and cell-derived xenograft mouse model were used to identify the biological function of RACK1. Flow cytometry and Seahorse assays were applied to examine cell cycle and oxygen consumption rate (OCR), respectively. Western blot, real-time PCR and autophagy double fluorescent assays were utilized to explore the signaling. Immunohistochemistry was performed to detect the expression of RACK1 and other indicators in tissue sections.
RESULTS RESULTS
Loss of RACK1 facilitated the viability, colony formation, cell cycle progression and OCR of GC cells in a glutamine-dependent manner. Further investigation revealed that RACK1 knockdown inhibited the lysosomal degradation of Alanine-serine-cysteine amino acid transporter 2 (ASCT2). Mechanistically, depletion of RACK1 remarkably decreased PTEN expression through up-regulating miR-146b-5p, leading to the activation of AKT/mTOR signaling pathway which dampened autophagy flux subsequently. Moreover, knockdown of ASCT2 could reverse the promotive effect of RACK1 depletion on GC tumor growth both in vitro and in vivo. Tissue microarray confirmed that RACK1 was negatively correlated with the expression of ASCT2 and p62, as well as the phosphorylation of mTOR.
CONCLUSION CONCLUSIONS
Together, our results demonstrate that the suppressive function of RACK1 in GC is associated with ASCT2-mediated glutamine metabolism, and imply that targeting RACK1/ASCT2 axis provides potential strategies for GC treatment.

Identifiants

pubmed: 37578594
doi: 10.1007/s13402-023-00854-1
pii: 10.1007/s13402-023-00854-1
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : National Natural Science Fund
ID : 82103535, 82273385, 82073245, 32271337
Organisme : National Natural Science Fund
ID : 82103535, 82273385, 82073245, 32271337
Organisme : National Natural Science Fund
ID : 82103535, 82273385, 82073245, 32271337

Informations de copyright

© 2023. Springer Nature Switzerland AG.

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Auteurs

Mengqian Chen (M)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Gaojia Wang (G)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Zhijian Xu (Z)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Jie Sun (J)

Department of General Surgery, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, People's Republic of China.

Bo Liu (B)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Lei Chang (L)

State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, People's Republic of China.

Jianxin Gu (J)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Yuanyuan Ruan (Y)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China.

Xiaodong Gao (X)

Department of General Surgery, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, People's Republic of China. xdgao_sh@hotmail.com.

Shushu Song (S)

Department of Biochemistry and Molecular Biology & NHC Key Laboratory of Glycoconjugates Research, School of Basic Medical Sciences, Fudan University, 138 Yixueyuan Road, Shanghai, 200032, People's Republic of China. shushusong@fudan.edu.cn.

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