title: [Role of PI3K/AKT signaling pathway in clonogenicity and tumorigenicity of CD90+ stem-like cells of the hepatocellular carcinoma cell line MHCC-97H].
abstract: OBJECTIVE
  To investigate the influence of the PI3K/AKT signaling pathway on the proportion and characteristics of the stem-like CD90(+) subpopulation of the human hepatocellular carcinoma (HCC) cell line MHCC-97.
  
  
  METHODS
  MHCC-97H cultures were treated with the PI3K/AKT pathway inhibitor LY294002. The proportion of the CD90(+) subpopulation was determined by flow cytometry, and the expression of related proteins was measured by Western blot. The clonogenicity of CD90(+) and CD90(-) cells was measured by plate colony formation assay. The tumorigenicity was compared between CD90(+) and CD90(-) subpopulations (with different concentrations) in xenograft experiments in nude mice, and the changes in tumorigenicity after the addition of LY294002 were evaluated. The changes in the expression of CD90, SHP2, P-AKT, and AKT in CD90(+) and CD90(-) cell xenografts after the addition of LY294002 were examined. Data were analyzed using t test.
  
  
  RESULTS
  LY294002 was capable of reducing the proportion of CD90(+) HCC stem cells from 2.98%±0.08% to 0.78%±0.08% (t = 32.400, P < 0.01) and reducing the clonogenicity of CD90(+) subpopulation from 95.13%±3.78% to 61.82%±7.23% (t = 7.617, P < 0.01). However, it showed no significant effect on the clonogenicity of CD90(-) subpopulation. LY294002 also reduced the tumorigenicity of CD90(+) subpopulation and the expression of CD90, SHP2, and P-AKT in related HCC stem cells, but it did not significantly affect the expression of AKT. LY294002 had no significant inhibitory effect on the tumorigenicity of CD90(-) cells.
  
  
  CONCLUSIONS
  The CD90(+) subpopulation of MHCC-97H cells has the characteristics of stem cells and is dependent on the PI3K/AKT signaling pathway.
authors.affiliation: Hepatobiliary Surgery Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China.
authors.forename: Y C
authors.initials: YC
authors.lastname: Peng
authors.forename: N F
authors.initials: NF
authors.lastname: Peng
authors.forename: Z B
authors.initials: ZB
authors.lastname: Xie
authors.forename: S D
authors.initials: SD
authors.lastname: Lu
authors.forename: J H
authors.initials: JH
authors.lastname: Zhong
authors.forename: Y B
authors.initials: YB
authors.lastname: Pang
authors.forename: S L
authors.initials: SL
authors.lastname: Zhu
authors.forename: X M
authors.initials: XM
authors.lastname: You
authors.forename: L Q
authors.initials: LQ
authors.lastname: Li
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D002867:Chromones; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009025:Morpholines; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000166:cytology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction; D018800:Thy-1 Antigens / Q000378:metabolism*
chemical_list: D002867:Chromones; D009025:Morpholines; D018800:Thy-1 Antigens; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt