Metabolomics analysis of the potential anticancer mechanism of annonaceous acetogenins on a multidrug resistant mammary adenocarcinoma cell

doi:10.1016/j.ab.2018.04.022

Analytical Biochemistry

Volume 553, 15 July 2018, Pages 1-6

https://doi.org/10.1016/j.ab.2018.04.022 Get rights and content

Highlights

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A metabolomics approach by UFLC-Q-TOF-MS coupled with pattern recognition methods to ACGs for its antitumor activity.
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Twenty three endogenous metabolites were identified and five metabolic pathways were found.
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Barely visible differences in the metabolites between ACGs types were demonstrated.

Abstract

Although annonaceous acetogenins (ACGs) have been reported to have antitumor activity for over three decades, and many of the underlying mechanism of ACGs on cancer have been clarified, there are still outstanding issues. In particular, the changes of small metabolite in cancer cells, caused by ACGs intake, have been reported rarely. Recent research has showed that cellular metabolic profiling coupled with ultra-flow liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UFLC-Q-TOF-MS) and multivariable statistical analysis enables a good understanding of ACGs' effects on multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells. As a result, 23 potential biomarkers (p < 0.05, VIP >1) were identified, and 5 pathways (impact-value > 0.10) identified. The differential metabolites suggested that ACGs affected metabolomics pathways, including arginine and proline metabolism, glycerophospholipid metabolism, taurine and hypotaurine metabolism, alanine, aspartate and glutamate metabolism and D-Glutamine and D-glutamate metabolism.

Graphical abstract

Introduction

Breast cancer is a major public health problem worldwide. World Cancer Report 2014 showed that the incidence rate and the mortality rate of breast cancer among women (43.3, 12.9 per 100,000) were much higher than that of any other cancers [1]. Multidrug resistance (MDR) is one of the major limitations on successful treatment for many cancers including breast cancer [2]. A cell's full complement of small-molecule metabolites, is a direct indicator of phenotypic diversity of single cells and nearly an immediate readout of how cells react to environmental influences [3]. It can give an insight into unknown processes in the MCF-7/Adr breast cancer cell after the treatment of annonaceous acetogenins.

Annonaceous acetogenins (ACGs), a series of polyketides found almost exclusively from Annonaceae plants, have been found to exhibit a broad range of biological properties, such as antineoplastic, antiparasitic, cytotoxic, immunosuppressive, neurotoxic and pesticidal effects [4]. According to their implicated structures, ACGs are divided into several classes, such as linear ACGs, epoxy ACGs, mono-THF ACGs, bis-THF ACGs (adjacent ACGs, non-adjacent ACGs), tri-THF ACGs and THP ACGs [5]. Recent research showed that bis-THF ACGs have a higher bioactivity than other types [6]. In our previous work, eight ACGs were isolated from Annona squamosa L., with two bis-THF ACGs showing significant toxicity against MDR cancer cell lines, including MCF-7/Adr breast cancer cells [7].

In this work, non-targeted metabolic profiling and multivariate statistical analysis were employed to investigate the changes of cellular metabolites after administrating two subtypes of bis-THF ACGs. A UFLC-QTOF/MS was used to identify metabolites in cells. Differential metabolites were revealed by multivariate statistical analysis in OPLS-DA (orthogonal partial least squares discriminate analysis) modes. The result shows that this two bis-THF ACGs regulate similar metabolic pathways in MCF-7/Adr cancer cells. It means that the changes of tetrahydrofuran ring moiety do not lead to the changes of underlying mechanism.

Section snippets

Chemicals and reagents

MS-grade acetonitrile was purchased from Tedia Company Inc (Darmstadt, Germany). Ultrapure water was prepared by a Mili-Q system from Millipore (Bedford, MA, USA). The chromatographically pure formic acid was purchased from Beijing Reagent Company (PR China). Squamocin P (M7) and Annosquatin III (M8) were isolated from Annona squamosa in our previous work, and the purity was over 98%. The IC₅₀ values of M7 and M8 were also reported as 3.34 and 4.04 μM [7].

Cell culture and treatments

The human breast cancer cell line

Metabolomics profiling

Intracellular metabolomics analysis was performed on a UFLC-QTOF-MS in negative and positive ion modes. PCA is an unsupervised method for visualizing the differences in metabolites among groups. It was applied here to visualize the score plats in negative and positive mode (Fig. 1). According to these figures, the bis-THF ACGs groups and the control group were distinguished clearly, indicating metabolic profiling changes after treating. Nevertheless, the M7H group, M7L group, M8H group and M8L

Conclusion

In this work, a metabolomics approach based on UFLC-Q-TOF-MS coupled with pathway analysis was developed and used to study the anticancer effects and mechanism of ACGs on MCF-7/Adr. As a result, 5 significant pathways were conducted, including arginine and proline metabolism, glycerophospholipid metabolism, taurine and hypotaurine metabolism, alanine, aspartate and glutamate metabolism and D-Glutamine and D-glutamate metabolism. These results provide an approach to understanding the key

Acknowledgements

This work was supported by the National Natural Science Foundation of China [grant number 81573577, 81403082 and 81274057].

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Metabolomics analysis of the potential anticancer mechanism of annonaceous acetogenins on a multidrug resistant mammary adenocarcinoma cell

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Chemicals and reagents

Cell culture and treatments

Metabolomics profiling

Conclusion

Acknowledgements

Colloids Surfaces B Biointerfaces

Nat. Prod. Res.

J. Nutr.

FEBS Lett.

J. Nutr.

J. Biol. Chem.

Trends Biochem. Sci.

Biochem. Biophys. Res. Commun.

Single-cell metabolomics: analytical and biological perspectives

Science