Expansion of CD133-expressing liver cancer stem cells in liver-specific phosphatase and tensin homolog deleted on chromosome 10-deleted mice

Stem Cells. 2009 Feb;27(2):290-9. doi: 10.1634/stemcells.2008-0332.

Abstract

PTEN (phosphatase and tensin homolog deleted on chromosome 10) is a lipid phosphatase that regulates mitogenic signaling pathways, and deficiency of PTEN results in cell proliferation, survival, and malignancy. Murine liver-specific Pten deletion models develop liver malignancy by 12 months of age. Using this model, we describe a population of CD133+ liver cancer stem cells isolated during the chronic injury phase of disease progression and before primary carcinoma formation. We performed immunohistochemistry and flow cytometry isolation using livers from 3- and 6-month-old Pten(loxP/loxP); Alb-Cre+ mice (mutants) and controls. CD133+CD45- nonparenchymal (NP) cells were analyzed for gene expression profile and protein levels. Single CD133+CD45- oval cells were isolated for clonal expansion and tumor analysis. Cultured and freshly isolated liver CD133+CD45- and CD133-CD45- NP cells were injected into immune-deficient and immune-competent mice. In mutant mice, the NP fraction increased in CD133+CD45- cells in 3- and 6-month-old Pten-deleted animals compared with controls. Clone lines expanded from single CD133+CD45- cells demonstrated consistent liver progenitor cell phenotype, with bilineage gene expression of hepatocyte and cholangiocyte markers. CD133+ cells from expanded clone lines formed robust tumors in immune-deficient and immune-competent mice. Furthermore, freshly isolated CD133+CD45- NP liver cells from 6-month-old mutants formed tumors in vivo, and CD133-CD45- NP cells did not. Consistent with a cancer stem cell phenotype, CD133+ cells demonstrate resistance to chemotherapy agents compared with CD133- cells. CD133+CD45- nonparenchymal cells from chronic injury Pten(loxP/loxP); Alb-Cre+ mice represent a bipotent liver progenitor cell population with cancer stem cell phenotype.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • AC133 Antigen
  • Animals
  • Antigens, CD / metabolism*
  • Blotting, Western
  • Cell Survival / genetics
  • Cell Survival / physiology
  • Cells, Cultured
  • Chromosomes, Mammalian / genetics*
  • Flow Cytometry
  • Glycoproteins / metabolism*
  • Immunohistochemistry
  • Leukocyte Common Antigens / metabolism
  • Liver Neoplasms / metabolism*
  • Liver Neoplasms / pathology*
  • Mice
  • Mice, Mutant Strains
  • Mice, Nude
  • Neoplastic Stem Cells / metabolism*
  • Neoplastic Stem Cells / pathology
  • PTEN Phosphohydrolase / genetics*
  • Peptides / metabolism*
  • Polymerase Chain Reaction

Substances

  • AC133 Antigen
  • Antigens, CD
  • Glycoproteins
  • Peptides
  • Prom1 protein, mouse
  • Leukocyte Common Antigens
  • PTEN Phosphohydrolase
  • Pten protein, mouse