Standardized flavonoid-rich fraction of Artemisia princeps Pampanini cv. Sajabal induces apoptosis via mitochondrial pathway in human cervical cancer HeLa cells

• Published in: Journal of ethnopharmacology 2012-05, Vol.141 (1), p.460-468
• Main Authors: Ju, Hye-Kyung, Lee, Heon-Woo, Chung, Kyung-Sook, Choi, Jung-Hye, Cho, Jin-Gyeong, Baek, Nam-In, Chung, Hae-Gon, Lee, Kyung-Tae
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Artemisia - chemistry; Artemisia princeps Pampanini cv. Sajabal; bcl-X Protein - genetics; bcl-X Protein - metabolism; Bcl-xL; Blotting, Western; Caspase Inhibitors; Caspases; Caspases - metabolism; Chemical Fractionation; Cysteine Proteinase Inhibitors; Cytochromes c - metabolism; Dose-Response Relationship, Drug; Enzyme Activation; Female; Flavonoids - isolation & purification; Flavonoids - pharmacology; HeLa Cells; Humans; Membrane Potential, Mitochondrial - drug effects; Mice; Mice, Inbred BALB C; Mice, Nude; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondria - pathology; Phytotherapy; Plant Components, Aerial; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Plants, Medicinal; Poly(ADP-ribose) Polymerases - metabolism; Time Factors; Transfection; Uterine Cervical Neoplasms - drug therapy; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - metabolism; Uterine Cervical Neoplasms - pathology; Xenograft Model Antitumor Assays
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2012.03.011

Our results indicate caspase-mediated activation of the mitochondrial death pathway plays a critical role in the FRAP-induced apoptosis of HeLa cells and that FRAP inhibits the in vivo tumor growth of HeLa xenograft mice. Artemisia princeps Pampanini is widely used in Eastern traditional medicine for the treatment of circulatory disorders, such as, dysmenorrhea, hematuria, hemorrhoids, and inflammation, and is also used to treat chronic conditions, such as, cancers, ulcers, and digestive disorders. The purpose of this study is to investigate the effect of a standardized flavonoid-rich fraction of Artemisia princeps Pampanini cv. Sajabal (FRAP) on the induction of apoptosis and the molecular mechanism involved in human cervical cancer HeLa cells. Human cervical cancer HeLa cells were treated with FRAP and apoptosis was detected by cell morphologic observation, annexin-V-PI staning and western blot analysis on the expression of protein associated with cell death. FRAP led to the cleavages of caspase-3, -8, and -9 and the cleavage of poly (ADP-ribose) polymerase (PARP) in HeLa cells. Caspase-3 inhibitor (z-DEVD-fmk), caspase-8 inhibitor (z-IETD-fmk), caspase-9 inhibitor (z-LEHD), and broad caspase inhibitor (z-VAD-fmk) significantly suppressed the FRAP-induced accumulation of annexin V positive cells. Furthermore, it was found that FRAP caused a loss of mitochondrial membrane potential (MMP) and the release of cytochrome c to the cytosol. Furthermore, the overexpression of Bcl-xL significantly prevented FRAP-induced apoptosis, MMP changes, and the activations of caspase-3, -8, and -9. Interestingly, pretreatment with caspase-8 inhibitor significantly reduced the FRAP-induced activation of caspase-3 but not that of caspase-9, whereas the caspase-3 inhibitor, z-DEVD-fmk, markedly attenuated the FRAP-induced activation of caspase-8. In BALB/cnu/nu mice bearing a HeLa xenograft, FRAP dosed at 25 or 50mg/kg significantly inhibited tumor growth. Our results indicate caspase-mediated activation of the mitochondrial death pathway plays a critical role in the FRAP-induced apoptosis of HeLa cells and that FRAP inhibits the in vivo tumor growth of HeLa xenograft mice.