Abstract

Metabolomic profiling is an increasingly important method for identifying potential biomarkers in cancer cells with a view towards improved diagnosis and treatment. Nuclear magnetic resonance (NMR) provides a potentially noninvasive means to accurately characterize differences in the metabolomic profiles of cells. In this work, we use 1H NMR to measure the metabolomic profiles of water soluble metabolites extracted from isogenic control and oncogenic HRAS-, KRAS-, and NRAS-transduced BEAS2B lung epithelial cells to determine the robustness of NMR metabolomic profiling in detecting differences between the transformed cells and their untransformed counterparts as well as differences among the RAS-transformed cells. Unique metabolomic signatures between control and RAS-transformed cell lines as well as among the three RAS isoform-transformed lines were found by applying principal component analysis to the NMR data. This study provides a proof of principle demonstration that NMR-based metabolomic profiling can robustly distinguish untransformed and RAS-transformed cells as well as cells transformed with different RAS oncogenic isoforms. Thus, our data may potentially provide new diagnostic signatures for RAS-transformed cells.

Highlights

  • It has long been appreciated that the metabolism of normal and malignant cells can significantly differ (Warburg, 1956)

  • One of the earliest applications of cellular Nuclear magnetic resonance (NMR) metabolomics has been to look for biomarkers associated with activation of the RAS oncogene (Aboagye & Bhujwalla, 1999; How to cite this article Marks et al (2016), 1H NMR studies distinguish the water soluble metabolomic profiles of untransformed and RAS-transformed cells

  • Due to the similar average values of the ‘‘effective’’ NMR glutamate content, xglutamate, observed in both the control and RAS-transformed cells (Fig. S1) and the relatively large glutamate signals observed in all cells lines, an alternative to the total intensity normalization scheme used in Eq (2) was investigated whereby the metabolite signals were normalized by the observed glutamate signal in each sample

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Summary

Introduction

It has long been appreciated that the metabolism of normal and malignant cells can significantly differ (Warburg, 1956). Due to the similar average values of the ‘‘effective’’ NMR glutamate content, xglutamate , observed in both the control and RAS-transformed cells (Fig. S1) and the relatively large glutamate signals observed in all cells lines (only the lactate signals were larger on average), an alternative to the total intensity normalization scheme used in Eq (2) was investigated whereby the metabolite signals were normalized by the observed glutamate signal in each sample.

Results
Conclusion
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