Regulatory role of hexosamine biosynthetic pathway on hepatic cancer stem cell marker CD133 under low glucose conditions.

Shu-Hai Lin,Li Fu,Zongwei Cai,Xiaoyan Ming,Tengfei Liu,Xin-Yuan Guan,Basem Kanawati,Zhi Tang,Philippe Schmitt-Kopplin

doi:10.1038/srep21184

Abstract

Cancer was hypothesized to be driven by cancer stem cells (CSCs), but the metabolic determinants of CSC-like phenotype still remain elusive. Here, we present that hexosamine biosynthetic pathway (HBP) at least in part rescues cancer cell fate with inactivation of glycolysis. Firstly, metabolomic analysis profiled cellular metabolome in CSCs of hepatocellular carcinoma using CD133 cell-surface marker. The metabolic signatures of CD133-positive subpopulation compared to CD133-negative cells highlighted HBP as one of the distinct metabolic pathways, prompting us to uncover the role of HBP in maintenance of CSC-like phenotype. To address this, CSC-like phenotypes and cell survival were investigated in cancer cells under low glucose conditions. As a result, HBP inhibitor azaserine reduced CD133-positive subpopulation and CD133 expression under high glucose condition. Furthermore, treatment of N-Acetylglucosamine in part restores CD133-positive subpopulation when either 2.5 mM glucose in culture media or glycolytic inhibitor 2-deoxy-D-glucose in HCC cell lines was applied, enhancing CD133 expression as well as promoting cancer cell survival. Together, HBP might be a key metabolic determinant in the functions of hepatic CSC marker CD133.

Highlights

The growing evidence elucidates some integral molecules and signaling pathways responsible for the stemness maintenance and differentiation of CD133-positive subpopulation cells[12,13,14], the metabolic features of CD133-positive enriched cancer stem cells (CSCs) and the effects of nutrients in CSCs remain inconclusive
We isolated CD133-positive and counterpart CD133-negative subsets from human liver cancer cell line PLC8024 followed by metabolomics study based on Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) and ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) to profile the cellular metabolome
Our metabolomic data raised a question whether hexosamine biosynthetic pathway (HBP) plays a key role for discriminating CD133-positive stemness phenotype in cancer cells

Summary

Introduction

The growing evidence elucidates some integral molecules and signaling pathways responsible for the stemness maintenance and differentiation of CD133-positive subpopulation cells[12,13,14], the metabolic features of CD133-positive enriched CSCs and the effects of nutrients in CSCs remain inconclusive. Metabolic rewiring in CSCs has not been well characterized. To address this issue, it would be necessary to perform the emerging technique metabolomics for cellular metabolome. We isolated CD133-positive and counterpart CD133-negative subsets from human liver cancer cell line PLC8024 followed by metabolomics study based on Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) and ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) to profile the cellular metabolome. Uridine diphosphate-N-acetylglucosamine (UDP-GlcNAc), an end product in hexosamine biosynthetic pathway (HBP) was determined as the elevated level in CD133-positive subsets. This finding prompted us to ask whether HBP impacts CSC-like phenotype. Our investigation demonstrated that HBP might coordinate with glycolytic pathway for the regulation of CD133 in hepatoma carcinoma cell lines

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Results

Conclusion