16-Hydroxycleroda-3,13-dien-15,16-olide Induces Apoptosis in Human Bladder Cancer Cells through Cell Cycle Arrest, Mitochondria ROS Overproduction, and Inactivation of EGFR-Related Signalling Pathways

• Published in: Molecules (Basel, Switzerland) Switzerland), 2020-08, Vol.25 (17), p.3958
• Main Authors: Chen, Yu-Chi, Wang, Po-Yu, Huang, Bu-Miin, Chen, Yu-Jen, Lee, Wei-Chang, Chen, Yung-Chia
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; Bcl-2 protein; Bladder cancer; c-Myc protein; Cancer; Cancer therapies; Caspase-3; Cell cycle; Cell Cycle Checkpoints - drug effects; Cell death; Cell division; Cell Line, Tumor; Cell proliferation; Chemical compounds; Chemotherapy; clerodane diterpene; Cyclin D1; Cyclin-dependent kinase inhibitor p27; Cyclin-dependent kinases; Cytochrome c; Cytotoxicity; Diterpenes; Diterpenes, Clerodane - pharmacology; Epidermal growth factor; epidermal growth factor receptor; Epidermal growth factor receptors; ErbB Receptors - metabolism; G1 phase; Genes; Humans; Hypoxia-inducible factor 1a; Inactivation; Intracellular signalling; Kinases; Membrane potential; Metastasis; Mitochondria; Mitochondria - drug effects; Myc protein; Oxidative stress; p53 Protein; Pharmacology; Poly(ADP-ribose) polymerase; Polyalthia - metabolism; Proteins; Reactive oxygen species; Reactive Oxygen Species - metabolism; Signal Transduction; Urinary Bladder Neoplasms - drug therapy; Urinary Bladder Neoplasms - pathology; Vascular endothelial growth factor
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules25173958

A clerodane diterpene compound 16-hydroxycleroda-3,13-dien-15,16-olide (CD) is considered a therapeutic agent with pharmacological activities. The present study investigated the mechanisms of CD-induced apoptosis in T24 human bladder cancer cells. CD inhibited cell proliferation in a concentration and time-dependent manner. CD-induced overproduction of reactive oxygen species and reduced mitochondrial membrane potential, associated with reduced expression of Bcl-2 and increased levels of cytosolic cytochrome c, cleaved PARP-1 and caspase-3. In addition, CD treatment led to cell cycle arrest at the G0/G1 phase and inhibited expression of cyclin D1 and cyclin-dependent kinases 2 and 4 and led to increased levels of p21, p27Kip1 and p53. All of these events were accompanied with a reduction of pEGFR, pMEK1/2, pERK1/2, pAkt, pmTOR, pP70S6K1, HIF-1α, c-Myc and VEGF. RNAseq-based analysis revealed that CD-induced cell death was characterised by an increased expression of stress and apoptotic-related genes as well as inhibition of the cell cycle-related genes. In summary, CD induces apoptosis in T24 bladder cancer cells through targeting multiple intracellular signaling pathways as a result of oxidative stress and cell cycle arrest.