Antitumor Effects of Esculetin, a Natural Coumarin Derivative, against Canine Mammary Gland Tumor Cells by Inducing Cell Cycle Arrest and Apoptosis

Abstract

Mammary gland tumors are the most common neoplasms in female dogs, of which 50% are malignant. Esculetin, a coumarin derivative, reportedly induces death in different types of cancer cells. In this study, we explore the anticancer effects of esculetin against CMT-U27 and CF41.mg canine mammary gland tumor cells. Esculetin significantly inhibited the viability and migration of both CMT-U27 and CF41.mg cells in a dose- and time-dependent manner. Flow cytometric analysis and terminal deoxynucleotidyl transferase dUTP nick-end labeling assay revealed increased numbers of annexin-V-positive cells and DNA fragmentation. Furthermore, a cell cycle analysis demonstrated that esculetin blocked the cell progression at the G0/G1 phase and the S phase in CMT-U27 and CF41.mg cells. These results were supported by a Western blot analysis, which revealed upregulated protein expression of cleaved caspase-3, a marker of apoptosis, and downregulated cyclin-dependent kinase 4 and cyclin D1 protein, the cell cycle regulators. In conclusion, this novel study proves that esculetin exerts in vitro antitumor effects by inducing apoptosis and cell cycle arrest in canine mammary gland tumors.

Esculetin unveiled: Decoding its anti‐tumor potential through molecular mechanisms—A comprehensive review

“…37 According to Choi et al CMT-U27, CF41.mg canine mammary gland tumor cells undergo apoptosis, and G0/G1 and S phase cell cycle arrest when exposed to esculetin at concentrations between 0 and 1 mM, and the expression of the cell death protein caspase-3 is upregulated by esculetin while the expression of the cell cycle regulators CDK4 and cyclin D1 is downregulated. 38 In conclusion, it is clear from the current mechanistic investigations of breast cancer that esculetin promotes the overexpression of tumor suppressor genes that can control cell cycle progression. This naturally occurring coumarin chemical also regulates the expression of numerous genes.…”

“…However, the bulk of these were ineffective for the intended use due to their numerous side effects. Esculetin alleviates drug resistance by downregulating cyclin D1 in breast cancer (CMT‐U27, CF41.mg), 38 colon cancer (HCT‐116 cell), 45 leukemia (HL‐60), 60 melanoma (G361 cell), 24 and renal cancer (786‐O and SN12‐PM6 cell) 26 . Esculetin dramatically reduced p65‐NF‐κB and ROS levels, improving anticancer drug sensitivity in resistant pancreatic cancer cell lines 18 .…”

“…Esculetin inhibited the growth of cancer cells, linoleic acid production, and cell proliferation when administered to MDA‐MB‐231 cancer cells at dosages of 40, 80, and 90 ng/mL 37 . According to Choi et al CMT‐U27, CF41.mg canine mammary gland tumor cells undergo apoptosis, and G0/G1 and S phase cell cycle arrest when exposed to esculetin at concentrations between 0 and 1 mM, and the expression of the cell death protein caspase‐3 is upregulated by esculetin while the expression of the cell cycle regulators CDK4 and cyclin D1 is downregulated 38 …”

Annulated Heterocyclic[g]Coumarin Composites: Synthetic Approaches and Bioactive Profiling

“…Using the coumarin as a backbone, Chen et al, [29] synthesized substituted furo[g]coumarins (6 and 7) and annulated a furan ring on it. They began their work using 1,2,3-benzenetriol (1) and used sulfuric acid as a catalyst to accomplish a Pechmann reaction with ethyl 3-oxobutyrate (2) to produce the substituted coumarin molecule (3). Compound (3) was then ether-linked with chloropropanone (4) to form compound (5) under the effect of propanone and K 2 CO 3 .…”

“…The development of effective and accessible pharmaceutical molecules to address a wide range of illnesses is a primary focus of medicinal chemistry. Because they are versatile chemical entities with antiproliferative, [1] antiradical, [2] antibacterial, [3] antifungal, [4] antiviral, [5,6] inflammatory suppressant, [7] hepatoprotective, [8] cardioprotective, [9] neuroprotective, [10] antidiabetic, [11] and lipid-lowering [12] properties, coumarin-based compounds have drawn a lot of attention in recent decades. Figure 1 illustrates the structure of coumarin, a dicyclic molecule that is made up of a five-membered lactone ring fused with an arene ring.…”