Study on the Intervention Mechanism of Cryptotanshinone on Human A2780 Ovarian Cancer Cell Line Using GC-MS-Based Cellular Metabolomics

Abstract

Cryptotanshinone (CT), an active component of the traditional Chinese medicine Salvia miltiorrhiza Bunge, exhibits a wide range of biological and pharmacological activities. Although the anticancer activity of CT is well known, the knowledge of its effect on the regulation of cancer cell metabolism is relatively new. The present study investigated the anticancer mechanism of CT in ovarian cancer with a focus on cancer metabolism. CCK8 assays, apoptosis assays, and cell cycle assays were conducted to reveal the growth-suppressive effect of CT on ovarian cancer A2780 cells. To explore the potential underlying mechanisms of CT, the changes in endogenous metabolites in A2780 cells before and after CT intervention were investigated using the gas chromatography–mass spectrometry (GC-MS) approach. A total of 28 important potential biomarkers underwent significant changes, mainly involving aminoacyl-tRNA biosynthesis, energy metabolism, and other pathways. Changes in the ATP and amino acid contents were verified with in vitro and in vivo experiments. Our results indicate that CT may exert an anti-ovarian cancer effect by inhibiting ATP production, promoting the protein catabolic process, and inhibiting protein synthesis, which may lead to cell cycle arrest and apoptosis.

Review on analytical technologies and applications in metabolomics

“…The preparation of cell samples must prevent further metabolism. During the analysis, cells were removed from the culture plate with a cell scraper [34,35]. The mixture was centrifuged (4000×g) for 30 min and the supernatant was sucked out.…”

“…Metabolomics originally used gas chromatography to analyze volatile components in plants [34,69,70]. As a result of the development of gas chromatography and mass spectrometry technology, it is commonly used gas chromatography and mass spectrometry analysis of common biological samples, such as urine, serum, plasma, bacteria, cells and so on [71][72][73].…”

“…Metabolites containing hydroxyl, amine, amide, phosphate and mercaptan groups can be derived by alkylmethylsilane method. Recently, many metabolomics studies of plant metabolites or urine samples have adopted GC-MS analysis techniques of methylsilylation derivatization or oximation and methylsilylation derivatization [34,76,79]. GC-MS, as a high-throughput metabolomics analysis method, can be used to quantify a large number of metabolites in a single assay [72, 80,81].…”