Artonin E Induces Apoptosis via Mitochondrial Dysregulation in SKOV-3 Ovarian Cancer Cells

• Published in: PloS one Vol. 11; no. 3; p. e0151466
• Main Authors: Rahman, Mashitoh Abd, Ramli, Faiqah, Karimian, Hamed, Dehghan, Firouzeh, Nordin, Noraziah, Ali, Hapipah Mohd, Mohan, Syam, Hashim, Najihah Mohd
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 28.03.2016; Public Library of Science (PLoS)
• Subjects: Acridine; Acridine orange; Activation; Analysis; Annexin V; Apoptosis; Apoptosis - drug effects; Apoptosis - genetics; Artocarpus; Artocarpus - chemistry; Artocarpus elasticus; Bark; Bcl-2 protein; Biology and Life Sciences; Blotting, Western; Cancer; Cancer therapies; Caspase 3 - metabolism; Caspase 8 - metabolism; Caspase 9 - metabolism; Cell cycle; Cell death; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Proliferation - genetics; Cell Survival - drug effects; Cell Survival - genetics; Complications and side effects; Cytochrome; Cytochrome c; Cytochromes c - metabolism; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA fragmentation; DNA Fragmentation - drug effects; Enzyme Activation - drug effects; Female; Flavonoids - chemistry; Flavonoids - pharmacology; Flow Cytometry; Heat shock proteins; Hsp70 protein; Humans; Influence; Inhibitory Concentration 50; Iodides; Medicine and Health Sciences; Membrane potential; Metastases; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Molecular Structure; Ovarian cancer; Ovarian carcinoma; Ovarian Neoplasms - genetics; Ovarian Neoplasms - metabolism; Ovarian Neoplasms - pathology; Oxygen; Pathways; Product development; Propidium iodide; Proteins; Proto-Oncogene Proteins c-bcl-2 - metabolism; Reactive oxygen species; Reactive Oxygen Species - metabolism; Research and analysis methods; S phase; S Phase Cell Cycle Checkpoints - drug effects; S Phase Cell Cycle Checkpoints - genetics; Signaling; Survivin; Toxicity; Malaysia
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0151466

Artonin E is a prenylated flavonoid isolated from the stem bark of Artocarpus elasticus Reinw.(Moraceae). This study aimed to investigate the apoptotic mechanisms induced by artonin E in a metastatic human ovarian cancer cell line SKOV-3 in vitro. MTT assay, clonogenic assay, acridine orange and propidium iodide double staining, cell cycle and annexin V analyses were performed to explore the mode of artonin E-induced cell death at different time points. DNA laddering, activation of caspases-3, -8, and -9, multi-parametric cytotoxicity-3 analysis by high-content screening, measurement of reactive oxygen species generation, and Western blot were employed to study the pathways involved in the apoptosis. MTT results showed that artonin E inhibited the growth of SKOV-3 cells, with IC50 values of 6.5±0.5 μg/mL after 72 h treatment, and showed less toxicity toward a normal human ovarian cell line T1074, with IC50 value of 32.5±0.5 μg/mL. Results showed that artonin E induced apoptosis and cell cycle arrest at the S phase. This compound also promoted the activation of caspases-3, -8, and -9. Further investigation into the depletion of mitochondrial membrane potential and release of cytochrome c revealed that artonin E treatment induced apoptosis via regulation of the expression of pro-survival and pro-apoptotic Bcl-2 family members. The expression levels of survivin and HSP70 proteins were also down regulated in SKOV-3 cells treated with artonin E. We propose that artonin E induced an antiproliferative effect that led to S phase cell cycle arrest and apoptosis through dysregulation of mitochondrial pathways, particularly the pro- and anti-apoptosis signaling pathways.