Journal: Journal of Cancer
Article Title: Molecular Mechanism of Cinnamomum verum Component Cuminaldehyde Inhibits Cell Growth and Induces Cell Death in Human Lung Squamous Cell Carcinoma NCI-H520 Cells In Vitro and In Vivo
Figure Lengend Snippet: Inhibition of NCI-H520 topoisomerase I and II activities by CuA. (A) CuA inhibited topoisomerase I activity in NCI-H520 cells. Nuclear proteins from NCI-H520 cells were added to a specific topoisomerase I reaction mixture in the presence of the indicated concentrations of CuA (lanes 3-5), or 60 μM CPT (lane 6), or the vehicle (1% DMSO, lane 2). Lane 1: pUC19 DNA only. (B) CuA inhibits topoisomerase II activity in NCI-H520 cells. DNA relaxation assay (upper part) and decatenation assay (lower part): Nuclear proteins from NCI-H520 cells were added to a specific topoisomerase II reaction mixture in the presence of the indicated concentrations of CuA (lanes 3-5) or 60 μM VP-16, a specific topoisomerase II inhibitor (lane 6), or the vehicle (1% DMSO, lane 2). (Lane 1) Supercoiled pUC19 DNA (upper part) or kDNA (lower part) only. kDNA is a large network of plasmids, and when it is analyzed by gel electrophoresis, it penetrates only slightly into agarose gel (result not shown). Upon decatenation by topoisomerase II, mini circles monomers of kDNA were formed (lower part, lanes 2-6). CPT, camptothecin; CuA, cuminaldehyde; S and R, supercoiled and the relaxed forms of the pUC 19 plasmid DNA, respectively; VP-16, etoposide.
Article Snippet: Cuminaldehyde (CuA) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich, Inc. (St. Louis, Mo, USA).
Techniques: Inhibition, Activity Assay, Cycling Probe Technology, Nucleic Acid Electrophoresis, Agarose Gel Electrophoresis, Plasmid Preparation