title: Extract of bulbus Fritillaria cirrhosa perturbs spindle assembly checkpoint, induces mitotic aberrations and genomic instability in human colon epithelial cell line.
abstract: BACKGROUND
  Bulbus Fritillaria cirrhosa D. Don (BFC) has been used in China as a folk medicine for the treatment of cough and asthma for more than 2000 years. The antitussive and antiasthmatic effects of BFC have been reported before, nevertheless its toxicity and safety have not been documented. This study investigated the possible effects of BFC on spindle assembly checkpoint (SAC), mitotic fidelity and genomic stability in human NCM460 colon epithelial cells.
  
  
  METHODS
  Cells were treated with BFC (0, 20, 40, 80 and 160μg/ml) for 24, 48 and 72h and harvested differently according to the biomarkers observed. Mitotic aberrations were assessed by the biomarkers of chromosome misalignment (CMA), chromosome lagging (CL) and chromatin bridge (CB). Frequencies of micronuclei (MN), nucleoplasmic bridge and nuclear bud (NB) in cytokinesis-block micronucleus assay were used as indicators of genomic instability (GIN). SAC activity was determined by anaphase to metaphase ratio (AMR) and the expression of several SAC genes, including CENP-E, Mps1, Bub1, Mad-1, BubR1 and Mad-2.
  
  
  RESULTS
  Compared with the control, cells in BFC treated groups (80 and 160μg/ml) showed: 1) increased AMR (p<0.05), up-regulated expression of Mps1, Bub1 and Mad-1 (p<0.05) and down-regulated expression of CENP-E, BubR1 and Mad-2 (p<0.05); 2) increased frequencies of CMA, CL and CB (p<0.01); 3) increased incidences of MN and NB (p<0.01).
  
  
  CONCLUSIONS
  This study revealed for the first time that BFC causes mitotic aberrations and GIN in human colon epithelial cells and these effects maybe the result of SAC dysfunction.
authors.affiliation: School of Life Sciences, The Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Yunnan Normal University, Kunming, Yunnan, China; School of Life Sciences, Yunnan University, Kunming, Yunnan, China.
authors.forename: Xihan
authors.initials: X
authors.lastname: Guo
authors.affiliation: School of Life Sciences, The Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Yunnan Normal University, Kunming, Yunnan, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Ni
authors.affiliation: Institute of Genetics, Fudan University, Shanghai, China.
authors.forename: Jinglun
authors.initials: J
authors.lastname: Xue
authors.affiliation: School of Life Sciences, The Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Yunnan Normal University, Kunming, Yunnan, China. Electronic address: wangxu@fudan.edu.cn.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
mesh_terms: D002460:Cell Line; D003106:Colon / Q000166:cytology / Q000187:drug effects*; D004365:Drugs, Chinese Herbal / Q000633:toxicity*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects*; D031410:Fritillaria / Q000737:chemistry* / Q000633:toxicity; D042822:Genomic Instability / Q000187:drug effects*; D006801:Humans; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects*; D008938:Mitosis / Q000187:drug effects*; D010936:Plant Extracts / Q000494:pharmacology; D018517:Plant Roots / Q000737:chemistry / Q000633:toxicity; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004365:Drugs, Chinese Herbal; D010936:Plant Extracts
keywords: Chromosome misalignment; Chromosome missegregation; Fritillaria cirrhosa; Micronucleus; Nuclear bud; Spindle assembly checkpoint