title: Novel insights into the apoptosis mechanism of DNA topoisomerase I inhibitor isoliquiritigenin on HCC tumor cell.
abstract: The inhibitory effect of DNA topoisomerase (Top I) by isoliquiritigenin(ISO) were investigated and their interaction mechanism was evaluated using methods including UV-vis absorption, fluorescence, coupled with molecular simulation, and using the MTT method of inhibition rate of HCC tumor cell SNU475 proliferation assay, finally, the interaction of ISO with calf thymus DNA was investigated by melting measurements and molecular docking studies. It was found that isoliquiritigenin reversibly inhibited DNA Top I in a competitive manner with the concentrations of ISO resulting in 50% activity lost (IC50) were estimated to be 0.178 ± 0.12 mM. Isoliquiritigenin exhibited a strong ability to quench the intrinsic fluorescence of Top I through a static quenching procedure. The positive values of enthalpy change and entropy change suggested that the binding of isoliquiritigenin to Top I was driven mainly by hydrophobic interactions. The molecular docking results revealed isoliquiritigenin actually interacted with the primary amino acid residues on the active site of Top I, and the detection results of fluorescence staining and the inhibitory effect on the growth of HCC SUN475 showed that isoliquiritigenin induced the apoptosis cells increased gradually. The interaction of ISO with DNA can cause the denaturation temperature to be increased, which indicated that the stabilization of the DNA helix was increased in the presence of ISO, which indicated that the results provide strong evidence for intercalative binding of ISO with DNA.
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China.
authors.forename: Ze-Xin
authors.initials: ZX
authors.lastname: Li
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China.
authors.forename: Kun
authors.initials: K
authors.lastname: You
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Xinxiang Medical University, 453100, China. Electronic address: wangjianguoxx@163.com.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wang
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D047188:Chalcones / Q000737:chemistry / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry; D006801:Humans; D062105:Molecular Docking Simulation; D059004:Topoisomerase I Inhibitors / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D047188:Chalcones; D059004:Topoisomerase I Inhibitors; D004247:DNA; C040920:isoliquiritigenin
keywords: Apoptosis cells; DNA topoisomerase I; Fluorescence quenching; Inhibition effect; Isoliquiritigenin; Melting measurements; Molecular simulation