Anticancer Activity and Cell Mechanism of a Novel Flavonoid Derivative from Chrysanthemum × morifolium (Ramat). Hemsl Leaves Against the MCF-7 Breast Cancer Line in vitro

Background: Breast cancer has the highest number of cases and deaths in women. Cell cycle dysregulation and decreased cell apoptosis are among the causes of cancer cell growth. An ideal anticancer drug has selective toxicity, meaning it can kill cancer cells without damaging normal tissue cells. Several cultivars of Chrysanthemum morifolium leaves (CML) contain the main active compounds, such as flavonoids, as active ingredients of drugs that have potential as anticancer agents. Purpose: The main objective of this study was to investigate the potential anticancer effects of a compound extracted from CML. Methods: Chromatographic isolation of compounds using different separation techniques, including extraction, separation, and purification; characterization using ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR), and mass spectroscopy; and evaluation of anticancer activity in vitro (WST-8 assay), cell mechanism: cell cycle inhibition, and apoptosis activity (flow cytometry), and analysis of the ESR1 gene expression (RT-qPCR). Results: Bioactive components found in the ethyl acetate extract of CML were acaetin ( 1 ), 3,5,5′-Trihydroxy-4′-methoxy flavone ( 2 ), and kaempferide-7-O-glycoside ( 3 ). The lowest percentage of cell viability was shown in isolate ( 2 ) with an IC 50 value of 49.37 and 225.045 ppm in MCF-7 and HEK-293 cells. Isolate 2 at a concentration of 1 × IC 50 caused a significant increase in the G2/M phase (50.73 ± 4.13%), indicating G2/M arrest. Meanwhile, in the apoptosis assay, the same treatment increased early apoptosis of up to 38.87 ± 11.43%. Isolate 2 CML showed increased ESR1 gene expression at a dose of 1 × IC 50 of 1.34 ± 0.13 and at a dose of ½ × IC 50 of 1.01 ± 0.14. Conclusion: Isolate 2 CML showed selective potential as an anticancer agent through G2/M cell cycle inhibition, early apoptosis, and suppression of ESR1 gene expression. Keywords: Chrysanthemum morifolium, cytotoxicity, MCF-7 cells, in vitro, cell mechanism