title: The crude extract of Corni Fructus induces apoptotic cell death through reactive oxygen species-modulated pathways in U-2 OS human osteosarcoma cells.
abstract: Crude extract of Corni Fructus (CECF) has been used in Traditional Chinese medicine for the treatment of different diseases for hundreds of years. The purpose of this study was to investigate the cytotoxic effects of CECF on U-2 OS human osteosarcoma cells. Flow cytometry was used for measuring the percentage of viable cells, cell-cycle distribution, apoptotic cells in sub-G1 phase, reactive oxygen species (ROS), Ca(2+) levels, and mitochondrial membrane potential (ΔΨm ). Comet assay and 4'-6-diamidino-2-phenylindole staining were used for examining DNA damage and condensation. Western blotting was used to examine apoptosis-associated protein levels in U-2 OS cells after exposed to CECF. Immunostaining and confocal laser system microscope were used to examine protein translocation after CECF incubation. CECF decreased the percentage of viability, induced DNA damage and DNA condensation, G₀/G₁ arrest, and apoptosis in U-2 OS cells. CECF-stimulated activities of caspase-8, caspase-9, and caspase-3, ROS, and Ca(2+) production, decreased ΔΨm levels of in U-2 OS cells. CECF increased protein levels of caspase-3, caspase-9, Bax, cytochrome c, GRP78, AIF, ATF-6α, Fas, TRAIL, p21, p27, and p16 which were associated with cell-cycle arrest and apoptosis. These findings suggest that CECF triggers apoptosis in U-2 OS cells via ROS-modulated caspase-dependent and -independent pathways.
authors.affiliation: Graduate Institute of Chinese Medicine, China Medical University, Taichung 404, Taiwan, Republic of China.
authors.forename: Ching-Lung
authors.initials: CL
authors.lastname: Liao
authors.forename: Shu-Chun
authors.initials: SC
authors.lastname: Hsu
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Yu
authors.forename: Jai-Sing
authors.initials: JS
authors.lastname: Yang
authors.forename: Nou-Ying
authors.initials: NY
authors.lastname: Tang
authors.forename: Wellington Gibson
authors.initials: WG
authors.lastname: Wood
authors.forename: Jaung-Geng
authors.initials: JG
authors.lastname: Lin
authors.forename: Jing-Gung
authors.initials: JG
authors.lastname: Chung
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001859:Bone Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D029768:Cornus / Q000737:chemistry*; D004247:DNA / Q000187:drug effects; D000091342:Endoplasmic Reticulum Chaperone BiP; D006801:Humans; D008516:Medicine, Chinese Traditional; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009363:Neoplasm Proteins / Q000378:metabolism; D012516:Osteosarcoma / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D010936:Plant Extracts / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D009363:Neoplasm Proteins; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D004247:DNA; D020169:Caspases