title: 8-bromo-7-methoxychrysin inhibits properties of liver cancer stem cells via downregulation of β-catenin.
abstract: OBJECTIVE
  To evaluate whether 8-bromo-7-methoxychrysin (BrMC), a synthetic analogue of chrysin, inhibits the properties of cancer stem cells derived from the human liver cancer MHCC97 cell line and to determine the potential mechanisms.
  
  
  METHODS
  CD133(+) cells were sorted from the MHCC97 cell line by magnetic activated cell sorting, and amplified in stem cell-conditioned medium to obtain the enriched CD133(+) sphere forming cells (SFCs). The stem cell properties of CD133(+) SFCs were validated by the tumorsphere formation assay in vitro and the xenograft nude mouse model in vivo, and termed liver cancer stem cells (LCSCs). The effects of BrMC on LCSCs in vitro were evaluated by MTT assay, tumorsphere formation assay and transwell chamber assay. The effects of BrMC on LCSCs in vivo were determined using a primary and secondary xenograft model in Balb/c-nu mice. Expressions of the stem cell markers, epithelial-mesenchymal transition (EMT) markers and β-catenin protein were analyzed by western blotting or immunohistochemical analysis.
  
  
  RESULTS
  CD133(+) SFCs exhibited stem-like cell properties of tumorsphere formation and tumorigenesis capacity in contrast to the parental MHCC97 cells. We found that BrMC preferentially inhibited proliferation and self-renewal of LCSCs (P < 0.05). Furthermore, BrMC significantly suppressed EMT and invasion of LCSCs. Moreover, BrMC could efficaciously eliminate LCSCs in vivo. Interestingly, we showed that BrMC decreased the expression of β-catenin in LCSCs. Silencing of β-catenin by small interfering RNA could synergize the inhibition of self-renewal of LCSCs induced by BrMC, while Wnt3a treatment antagonized the inhibitory effects of BrMC.
  
  
  CONCLUSIONS
  BrMC can inhibit the functions and characteristics of LCSCs derived from the liver cancer MHCC97 cell line through downregulation of β-catenin expression.
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Mei-Fang
authors.initials: MF
authors.lastname: Quan
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Xiao
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Liu
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Guo
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Kai-Qun
authors.initials: KQ
authors.lastname: Ren
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: Cao
authors.affiliation: Mei-Fang Quan, Li-Hong Xiao, Kai-Qun Ren, Fei Liu, Jian-Guo Cao, Xi-Yun Deng, Medical College, Hunan Normal University, Changsha 410013, Hunan Province, China.
authors.forename: Xi-Yun
authors.initials: XY
authors.lastname: Deng
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005419:Flavonoids / Q000494:pharmacology*; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000378:metabolism; D034622:RNA Interference; D018874:Spheroids, Cellular; D013997:Time Factors; D014162:Transfection; D047368:Tumor Burden / Q000187:drug effects; D060449:Wnt Signaling Pathway / Q000187:drug effects; D060509:Wnt3A Protein / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: C552860:8-bromo-7-methoxychrysin; D000071916:AC133 Antigen; D015703:Antigens, CD; C495270:CTNNB1 protein, human; D017077:Culture Media, Conditioned; D005419:Flavonoids; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; C555510:WNT3A protein, human; D060509:Wnt3A Protein; D051176:beta Catenin
keywords: 8-bromo-7-methoxychrysin; Cancer stem cell; Liver cancer; Self-renewal; β-catenin