title: Millepachine, a novel chalcone, induces G2/M arrest by inhibiting CDK1 activity and causing apoptosis via ROS-mitochondrial apoptotic pathway in human hepatocarcinoma cells in vitro and in vivo.
abstract: In this study, we reported millepachine (MIL), a novel chalcone compound for the first time isolated from Millettia pachycarpa Benth (Leguminosae), induced cell cycle arrest and apoptosis in human hepatocarcinoma cells in vitro and in vivo. In in vitro screening experiments, MIL showed strong antiproliferation activity in several human cancer cell lines, especially in HepG2 cells with an IC50 of 1.51 µM. Therefore, we chose HepG2 and SK-HEP-1 cells to study MIL's antitumor mechanism. Flow cytometry showed that MIL induced a G2/M arrest and apoptosis in a dose-dependent manner. Western blot demonstrated that MIL-induced G2/M arrest was correlated with the inhibition of cyclin-dependent kinase 1 activity, including a remarkable decrease in cell division cycle (cdc) 2 synthesis, the accumulation of phosphorylated-Thr14 and decrease of phosphorylation at Thr161 of cdc2. This effect was associated with the downregulation of cdc25C and upmodulation of checkpoint kinase 2 in response to DNA damage. MIL also activated caspase 9 and caspase 3, and significantly increased the ratio of Bax/Bcl-2 and stimulated the release of cytochrome c into cytosol, suggesting MIL induced apoptosis via mitochondrial apoptotic pathway. Associated with those effects, MIL also induced the generation of reactive oxygen species. In HepG2 tumor-bearing mice models, MIL remarkably and dose dependently inhibited tumor growth. Treatment of mice with MIL (20mg/kg intravenous [i.v.]) caused more than 65% tumor inhibition without cardiac damage compared with 47.57% tumor reduction by 5mg/kg i.v. doxorubicin with significant cardiac damage. These effects suggested that MIL and its easily modified structural derivative might be a potential lead compound for antitumor drug.
authors.affiliation: State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Keyuan Road 4, Gaopeng Street, Chengdu 610041, China.
authors.forename: Wenshuang
authors.initials: W
authors.lastname: Wu
authors.forename: Haoyu
authors.initials: H
authors.lastname: Ye
authors.forename: Li
authors.initials: L
authors.lastname: Wan
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Han
authors.forename: Guangcheng
authors.initials: G
authors.lastname: Wang
authors.forename: Jia
authors.initials: J
authors.lastname: Hu
authors.forename: Minhai
authors.initials: M
authors.lastname: Tang
authors.forename: Xingmei
authors.initials: X
authors.lastname: Duan
authors.forename: Yi
authors.initials: Y
authors.lastname: Fan
authors.forename: Shichao
authors.initials: S
authors.lastname: He
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.forename: Heying
authors.initials: H
authors.lastname: Pei
authors.forename: Xuewei
authors.initials: X
authors.lastname: Wang
authors.forename: Xiuxia
authors.initials: X
authors.lastname: Li
authors.forename: Caifeng
authors.initials: C
authors.lastname: Xie
authors.forename: Ronghong
authors.initials: R
authors.lastname: Zhang
authors.forename: Zhu
authors.initials: Z
authors.lastname: Yuan
authors.forename: Yongqiu
authors.initials: Y
authors.lastname: Mao
authors.forename: Yuquan
authors.initials: Y
authors.lastname: Wei
authors.forename: Lijuan
authors.initials: L
authors.lastname: Chen
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis; D016203:CDC2 Protein Kinase / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D002599:Chalcone / Q000494:pharmacology; D047188:Chalcones / Q000737:chemistry / Q000494:pharmacology*; D019926:Cyclin B / Q000235:genetics / Q000378:metabolism; D018844:Cyclin-Dependent Kinases; D003600:Cytosol / Q000378:metabolism; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005260:Female; D005434:Flow Cytometry; D059565:G2 Phase Cell Cycle Checkpoints; D056945:Hep G2 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008113:Liver Neoplasms / Q000473:pathology; D008564:Membrane Potentials; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D033641:Millettia / Q000737:chemistry; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D047188:Chalcones; D019926:Cyclin B; D017382:Reactive Oxygen Species; C583284:millepachine; D002599:Chalcone; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; D018844:Cyclin-Dependent Kinases