Cell metabolomics to study the function mechanism of Cyperus rotundus L. on triple-negative breast cancer cells

Abstract

Background: Triple-negative breast cancer (TNBC) is a kind of malignant tumor with higher recurrence and metastasis rate. According to historical records, the dry rhizomes Cyperus rotundus L. could be ground into powder and mixed with ginger juice and wine for external application for breast cancer. We studied the effect of the ethanol extract of Cyperus rotundus L. (EECR) on TNBC cells and found its' apoptosis-inducing effect with a doserelationship. But the function mechanism of EECR on TNBC is still mysterious. Hence, the present research aimed to detect its function mechanism at the small molecule level through ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) metabolomics. Methods: The CCK-8 assay and the Annexin V-FITC/PI assay were applied to test the effect of EECR on MDA-MB-231 cells and MDA-MB 468 cells at various concentrations of 0, 200, 400, and 600 μg/ml. UPLC-Q-TOF-MS/MS based metabolomics was used between the control group and the EECR treatment groups. Multivariate statistical analysis was used to visualize the apoptosis-inducing action of EECR and filtrate significantly changed metabolites. Results: The apoptosis-inducing action was confirmed and forty-nine significantly changed metabolites (VIP > 1, p < 0.05, and FC > 1.2 or FC < 0.8) were identified after the interference of EECR. The level of significant differential metabolites between control group, middle dose group, and high dose group were compared and found that which supported the apoptosis-inducing action with dose-dependence. Conclusion: By means of metabolism, we have detected the mechanism of EECR inducing apoptosis of TNBC cells at the level of small molecule metabolites and found that EECR impacted the energy metabolism of TNBC cells. In addition, we concluded that EECR induced apoptosis by breaking the balance between ATP-production and ATP-consumption: arresting the pathways of Carbohydrate metabolism such as Central carbon metabolism in cancer, aerobic glycolysis, and Amino sugar and nucleotide sugar metabolism, whereas accelerating the pathways of ATP-consumption including Amino Acids metabolism, Fatty acid metabolism, Riboflavin metabolism and Purine metabolism. Although further study is still needed, EECR has great potential in the clinical treatment of TNBC with fewer toxic and side effects.

The Natural Products and Extracts: Anti‐Triple‐Negative Breast Cancer in Vitro

“…[83] Besides, Yu et al adopted cell metabolomics to study the anticancer mechanism that EECR induced apoptosis by breaking the balance between ATP-production and ATP-consumption. [84] The lycopene-rich extracts obtained from red guava fruit can induce apoptosis in estrogen receptorpositive breast cancers. Yu et al studied the anticancer activity of two extract types from red guava fruit, total extracts, and smaller molecular weight (< 30 kDa) extracts (LMW extracts).…”

“…When the autophagy inhibitor 3‐methyladenine (3‐MA) was treated in TNBC cells, it could increase EECR sensitivity [83] . Besides, Yu et al adopted cell metabolomics to study the anticancer mechanism that EECR induced apoptosis by breaking the balance between ATP‐production and ATP‐consumption [84] …”

Unravelling the Anticancer Mechanisms of Traditional Herbal Medicines with Metabolomics

“…With the aid of metabolomics, a study recently reported that Cyperus rotundus likely induced apoptosis in TNBC cells by causing an arrest of aerobic glycolysis and increasing the pathways of ATP-consumption like amino acids metabolism, fatty acid metabolism, riboflavin metabolism, and purine metabolism, consequently leading to ATP depletion and cell death [134]. These altered metabolic profiles can serve as a basis for further hypothesis formulation in mechanistic follow-up studies to assess its effects on some key players (enzymes, protein) in these metabolic pathways.…”

Protective Mechanism of Gandou Decoction in a Copper-Laden Hepatolenticular Degeneration Model: In Vitro Pharmacology and Cell Metabolomics

“…Compared with the tissues or organisms, cell metabolomics has obvious advantages such as lower costs, better stability and reproducibility, fewer ethical issues, among others ( Chrysanthopoulos et al, 2010 ; Cuperlović-Culf et al, 2010 ). In the past few years, cell metabolomics has been used for revealing the therapeutic mechanisms underlying the use of herbs alone or TCM formulae in diseases ( Sun et al, 2018 ; Zhang et al, 2019 ; Ma et al, 2020 ). The major analytical techniques in metabolomics are nuclear magnetic resonance (NMR) spectroscopy, gas chromatography–mass spectrometry (GC-MS), capillary electrophoresis–mass spectrometry (CE-MS), and liquid chromatography–mass spectrometry (LC-MS).…”