Hinokitiol Exhibits Antitumor Properties through Induction of ROS-Mediated Apoptosis and p53-Driven Cell-Cycle Arrest in Endometrial Cancer Cell Lines (Ishikawa, HEC-1A, KLE)

Abstract

Hinokitiol is a natural tropolone derivative that is present in the heartwood of cupressaceous plants, and has been extensively investigated for its anti-inflammatory, antioxidant, and antitumor properties in the context of various diseases. To date, the effects of hinokitiol on endometrial cancer (EC) has not been explored. The purpose of our study was to investigate the anti-proliferative effects of hinokitiol on EC cells. Cell viability was determined with an MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and the quantification of apoptosis and reactive oxygen species (ROSs) was performed by using flow cytometry, while protein expression was measured with the Western blotting technique. Hinokitiol significantly suppressed cell proliferation through the inhibition of the expression of cell-cycle mediators, such as cyclin D1 and cyclin-dependent kinase 4 (CDK4), as well as the induction of the tumor suppressor protein p53. In addition, hinokitiol increased the number of apoptotic cells and increased the protein expression of cleaved-poly-ADP-ribose polymerase (PARP) and active cleaved-caspase-3, as well as the ratio of Bcl-2-associated X protein (Bax) to B-cell lymphoma 2 (Bcl-2). Interestingly, except for KLE cells, hinokitiol induced autophagy by promoting the accumulation of the microtubule-associated protein light chain 3B (LC3B) and reducing the sequestosome-1 (p62/SQSTM1) protein level. Furthermore, hinokitiol triggered ROS production and upregulated the phosphorylation of extracellular-signal-regulated kinase (p-ERK1/2) in EC cells. These results demonstrate that hinokitiol has potential anti-proliferative and pro-apoptotic benefits in the treatment of endometrial cancer cell lines (Ishikawa, HEC-1A, and KLE).

One-pot synthesis of cyclic-aminotropiminium carboxylate derivatives with DNA binding and anticancer properties

“…Our result of cell viability depicts that 6j and 6n are less toxic to HEK293 compared to HeLa cells, which is similar to observations made by treatment with natural compounds β-Thujaplicin in breast cancer cells, colon cancer and OSF 52 . Upon treatment of HeLa cells with our compounds (6e, 6j, and 6n) we detected an increase in proapoptotic markers like P53 and P38 which is in accordance with the other studies with natural tropolone derivatives [53][54][55] . Similarly, synthetic derivative alpha naphthyl tropolone induced P53/p-mTOR/p-AKT signaling which increased caspase 3/7 activity in leukemia cells 11 .…”

Anti-tumor pharmacology of natural products targeting mitosis

“…In recent years, hinokitiol has been found to have notable anti-tumor activity, including inhibiting tumor cell migration, promoting cell apoptosis, and inducing autophagy, thus highlighting its great potential for treating tumors. Hinokitiol treatment of endometrial carcinoid cells increases p53, decreases cyclin D1 and CDK4, and blocks cell growth in G0/G1 phase 139 . In human hepatocellular carcinoma cells, hinobinol mainly causes a decrease in CDK7, cyclin D1, and cyclin A2, thus blocking the S phase 140 .…”

Natural Kinase Inhibitors for the Treatment and Management of Endometrial/Uterine Cancer: Preclinical to Clinical Studies

“…Hinokitiol showed significant anti-proliferative activity against human endometrial cancer cell lines (Ishikawa, HEC-1A, and KLE). The IC 50 value was 13.33 μM in the Ishikawa cells, 49.51 μM in the HEC-1A cells, and 4.69 μM in the KLE cells ( Chen et al, 2021 ). Zhang and the team had studied the antiproliferative effects of garcinol against endometrial cancer cells (Ishikawa (ISH) and HEC-1B cell lines).…”