Induction of mitotic arrest and apoptosis by a novel synthetic quinolone analogue, CWC-8, via intrinsic and extrinsic apoptotic pathways in human osteogenic sarcoma U-2 OS cells.
CWC-8 is a new synthesized novel 2-phenyl-4-quinolone compound in our laboratory which has demonstrated potential antitumor activity. In this study, we have defined the viability inhibition and apoptotic mechanisms of CWC-8 on human osteogenic sarcoma U-2 OS cells. According to the MTT assay, the cell viability was inhibited by CWC-8 in a dose- and time-dependent manner, with an IC(50) of 4.97+/-0.24 microM. CWC-8 treatment induced G(2)/M arrest and apoptosis in U-2 OS cells by cell cycle and flow cytometry analysis. It also profoundly caused a decrease in polymerized tubulin levels by in vitro tubulin polymerization assay which indicated that the microtubular cytoskeleton appears to be one of the cellular targets in response to CWC-8. Western blotting and CDK1 kinase assay showed that CWC-8 treatment caused a time-dependent increase of Cyclin B and CDK1 protein levels and activity during G(2)/M arrest. CWC-8 treatment also caused a time-dependent increase in Fas/CD95, FADD, cytosolic cytochrome c, caspase-8/-9/-3 active form, Apaf-1, AIF, Bax protein levels, and decrease in Bcl-2 protein level. CWC-8 also promoted caspase-8/-9 and -3 activities; however, pretreatment of cells with pan-caspase, caspase-8/-9 and -3 inhibitors led to reduced cell growth inhibition action. Taken together, these findings show CWC-8 could be a potential candidate for cancer therapy.
Chang
Yu-Hsun
YH
Graduate Institute of Pharmaceutical Chemistry, China Medical University, Taichung 404, Taiwan, R.O.C.
Yang
Jai-Sing
JS
Kuo
Sheng-Chu
SC
Chung
Jing-Gung
JG
D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D004149:Dioxoles / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008938:Mitosis / Q000187:drug effects*; D012516:Osteosarcoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D015363:Quinolones / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D014407:Tumor Cells, Cultured
C543890:6-(naphthalen-1-yl)(1,3)dioxolo(4,5-g)quinolin-8(5H)-one; D000970:Antineoplastic Agents; D004149:Dioxoles; D015363:Quinolones; D016203:CDC2 Protein Kinase; D020169:Caspases