Carnosine suppresses human glioma cells under normoxic and hypoxic conditions partly via inhibiting glutamine metabolism

Acta Pharmacologica Sinica (2020)Cite this article

Abstract

L-Carnosine (β-alanyl-L-histidine) is a naturally occurring dipeptide, which has shown broad-spectrum anticancer activity. But the anticancer mechanisms and regulators remain unknown. In this study, we investigated the effects of carnosine on human glioma U87 and U251 cell lines under normoxia (21% O2) and hypoxia (1% O2). We showed that carnosine (25−75 mM) dose-dependently inhibited the proliferation of the glioma cells; carnosine (50 mM) inhibited their colony formation, migration, and invasion capacity. But there was no significant difference in the inhibitory effects of carnosine under normoxia and hypoxia. Treatment with carnosine (50 mM) significantly decreased the expression of glutamine synthetase (GS) at the translation level rather than the transcription level in U87 and U251 cells, both under normoxia and hypoxia. Furthermore, the silencing of GS gene with shRNA and glutamine (Gln) deprivation significantly suppressed the growth, migratory, and invasive potential of the glioma cells. The inhibitory effect of carnosine on U87 and U251 cells was partly achieved by inhibiting the Gln metabolism pathway. Carnosine reduced the expression of GS in U87 and U251 cells by promoting the degradation of GS through the proteasome pathway, shortening the protein half-life, and reducing its stability. Given that targeting tumor metabolism is a proven efficient therapeutic tactic, our results may present new treatment strategies and drugs for improving the prognosis of gliomas.

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Fig. 1: Effects of carnosine on the viability, colony formation, and apoptosis of U87 and U251 cells cultured under normoxic and hypoxic conditions.
Fig. 2: Carnosine inhibited U87 and U251 cell migration and invasion under normoxia and hypoxia.
Fig. 3: Carnosine downregulated GS at only the protein expression but did not affect its RNA expression.
Fig. 4: Time- and concentration-dependent inhibitory effect of carnosine on GS expression in U87 and U251 cells under conditions of normoxia, hypoxia, Gln (4 mM) treatment, and Gln deprivation.
Fig. 5: Carnosine combined with Gln deprivation inhibited the growth of U87 and U251 cells and induced apoptosis.
Fig. 6: The inhibitory effect of carnosine on the proliferation and metastasis of U87 and U251 cells was partly achieved by inhibiting Gln metabolism.
Fig. 7: Carnosine reduced the stability of the GS protein in U87 and U251 cells, shortened the half-life of the GS protein, and accelerated degradation of the GS protein by the proteasome pathway.

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Acknowledgements

This work was supported by Zhejiang Provincial Scientific Research Foundations (LY19H090010), Wenzhou City Science and Technology Project (Y20170014), and Key Discipline of Zhejiang Province in Medical Technology (First Class, Category A).

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Affiliations

  1. Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Yu-jia Fang, Ming Wu, Hai-ni Chen, Tian-tian Wen, Jian-xin Lyu & Yao Shen

  2. Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, Hangzhou, 310014, China

    Jian-xin Lyu

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  1. Yu-jia Fang
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  2. Ming Wu
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  3. Hai-ni Chen
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Contributions

YS and JXL conceived and designed the experiments; YJF and YS wrote the paper; YJF, MW, HNC, and TTW performed experiments and analyzed the data.

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Correspondence to Yao Shen.

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Fang, Yj., Wu, M., Chen, Hn. et al. Carnosine suppresses human glioma cells under normoxic and hypoxic conditions partly via inhibiting glutamine metabolism. Acta Pharmacol Sin (2020). https://doi.org/10.1038/s41401-020-0488-1

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Keywords

  • glioma
  • hypoxia
  • carnosine
  • glutamine synthetase
  • proteasome
  • tumor metabolism

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