title: Mangiferin induces cell cycle arrest at G2/M phase through ATR-Chk1 pathway in HL-60 leukemia cells.
abstract: This study aimed to determine the effect of mangiferin on the cell cycle in HL-60 leukemia cells and expression of the cell cycle-regulatory genes Wee1, Chk1 and CDC25C and to further investigate the molecular mechanisms of the antileukemic action of mangiferin. The inhibitory effect of mangiferin on HL-60 leukemia cell proliferation was determined by the MTT assay. The impact of mangiferin on the HL-60 cell cycle was evaluated by flow cytometry. After the cells were treated with different concentrations of mangiferin, the expression levels of Wee1, Chk1 and CDC25C mRNA were determined by RT-PCR, and Western blot was used to evaluate the expression levels of cdc25c, cyclin B1, and Akt proteins. The inhibition of HL-60 cell growth by mangiferin was dose- and time-dependent. After treatment for 24 h, cells in G2/M phase increased, and G2/M phase arrest appeared with increased mRNA expression of Wee1, Chk1 and CDC25C. Mangiferin inhibited Chk1 and cdc25c mRNA expression at high concentrations and induced Wee1 mRNA expression in a dose-dependent manner. It significantly inhibited ATR, Chk1, Wee1, Akt, and ERK1/2 phosphorylation but increased cdc2 and cyclin B1 phosphorylation. Furthermore, mangiferin reduced cdc25c, cyclin B1, and Akt protein levels while inducing Wee1 protein expression. It also antagonized the phosphorylation effect of vanadate on ATR, and the phosphorylation effect of EGF on Wee1. These findings indicated that mangiferin inhibits cell cycle progression through the ATR-Chk1 stress response DNA damage pathway, leading to cell cycle arrest at G2/M phase in leukemia cells.
authors.affiliation: Department of Hematology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China zhigangpengcn@126.com.
authors.forename: Z G
authors.initials: ZG
authors.lastname: Peng
authors.affiliation: Department of Hematology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China.
authors.forename: Y B
authors.initials: YB
authors.lastname: Yao
authors.affiliation: Department of Hematology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China.
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Hematology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China.
authors.forename: Y L
authors.initials: YL
authors.lastname: Tang
authors.affiliation: Department of Hematology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China.
authors.forename: X
authors.initials: X
authors.lastname: Huang
mesh_terms: D064007:Ataxia Telangiectasia Mutated Proteins / Q000096:biosynthesis / Q000235:genetics; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D015973:Gene Expression Regulation, Leukemic; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D009363:Neoplasm Proteins / Q000096:biosynthesis; D011494:Protein Kinases / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger; D015398:Signal Transduction / Q000187:drug effects; D044004:Xanthones / Q000008:administration & dosage*
chemical_list: D009363:Neoplasm Proteins; D012333:RNA, Messenger; D044004:Xanthones; C013592:mangiferin; D011494:Protein Kinases; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1