Costunolide Inhibits the Growth of OAW42-A Multidrug-Resistant Human Ovarian Cancer Cells by Activating Apoptotic and Autophagic Pathways, Production of Reactive Oxygen Species (ROS), Cleaved Caspase-3 and Cleaved Caspase-9

Abstract

Background Worldwide, ovarian cancer has a high mortality rate due to the difficulty in diagnosing early-stage disease and resistance to chemotherapy agents. Costunolide is a plant-derived sesquiterpene lactone with anti-oxidant properties. This study aimed to investigate the effects of costunolide on cell growth, apoptosis, autophagy, the production of reactive oxygen species (ROS), cleaved caspase-3, and cleaved caspase-9 on the multidrug-resistant ovarian cancer cell line, OAW42-A. Material/Methods The MTT assay determined the proliferation rate of OAW42-A multidrug-resistant ovarian cancer cells and the apoptosis rate was determined using propidium iodide (PI) staining. Autophagy was detected by measuring the expression of LC3 II. Fluorescence flow cytometry was used to measure the levels of reactive oxygen species (ROS) and the mitochondrial membrane potential. Protein expression of LC3 II, beclin 1, cleaved caspase-3, and cleaved caspase-9 were measured by Western blot. Results Costunolide treatment inhibited the growth of OAW42-A cells with an IC 50 of 25 μM, resulted in apoptotic cell death, increased the expression of Bax, and decreased the expression of Bcl-2. Confocal electron microscopy showed that costunolide induced autophagy in the OAW42-A cells. Western blot showed that costunolide treatment of OAW42-A cells increased the expression of the LC3 II, beclin 1, cleaved caspase-3, and cleaved caspase-9. Costunolide treatment significantly increased the levels of ROS and reduced the OAW42-A cell mitochondrial membrane potential. Conclusions Costunolide inhibited growth, apoptosis, ROS generation, and was associated with loss of mitochondrial membrane potential of OAW42-A multidrug-resistant ovarian cancer cells.

Triptolide inhibits JAK2/STAT3 signaling and induces lethal autophagy through ROS generation in cisplatin‑resistant SKOV3/DDP ovarian cancer cells

“…Consistent with our hypothesis, pretreatment with the antioxidant NAC substantially inhibited ROS generation and rescued TPL-induced autophagy, as supported by our results revealing that NAC markedly blocked TPL-induced autophagosome/autolysosome formation, LC3 conversion and p62 degradation in SKOV3/DDP cells. These data were consistent with those of previous studies which revealed that costunolide and cinobufagin induced autophagy in cancer cells through ROS production ( 33 , 44 ).…”

Costunolide—A Bioactive Sesquiterpene Lactone with Diverse Therapeutic Potential

“…Furthermore, incubation of multidrug resistant ovarian cancer cells (OAW42-A) with costunolide attenuated cell growth with an IC 50 of 25 µM, and induced apoptosis, which was mediated through induction of Bax, decreased the expression of Bcl-2 and cleavage of caspase-3 and 9. Moreover, the compound induced autophagy as evidenced by the elevated expression of LC3 II and Beclin 1 [65].…”

Undifferentiated Pleomorphic Sarcoma With Hyaline Globules (Thanatosomes)

“…Cleaved caspase 3 functions as a kind of control tower of apoptosis: it cleaves poly (ADP-ribose) polymerase (PARP), cytokeratin 18, vimentin, actin and other intracytoplasmic proteins. Cleaved caspase 3 has been applied to detecting apoptotic neoplastic cells in paraffin sections [ 24 - 26 ].…”