Dysplasia of human prostate CD133(hi) sub-population in NOD-SCIDS is blocked by c-myc anti-sense

Prostate. 2009 May 15;69(7):689-98. doi: 10.1002/pros.20918.

Abstract

Background: The CD133(hi) sub-population of prostate epithelial cells has been demonstrated to possess tumor-initiating capacity consistent with that of the cancer stem cell theory. However, the involvement of oncogenes such as c-myc has not been fully elucidated in the CD133(hi) sub-population.

Methods: We have isolated primary prostate cell strains (IBC-10a) and immortalized them by transfection with hTERT. The in vitro and in vivo tumorigenic capacity of isolated CD133(hi) and CD133(lo) cells was evaluated with respect to c-myc expression using specific sense and anti-sense oligonucleotides.

Results: Freshly immortalized cells consisted of <3.3% CD133(hi)/CD24(hi) sub-population (SP). "Prostaspheres" generated from single CD133(hi) cells in the presence of EGF consisted of approximately 10% CD133(hi) SPs in 12-21 day cultures. A single Prostasphere generated from single CD133(hi) cells (6-10 cell stage at day 6 injected i.t.) produced dysplastic lesions in NOD-SCID mice (n = 4/5). Treatment of Prostaspheres from CD133(hi) SPs in vitro with c-myc or cyclin D1 anti-sense oligonucleotides totally blocked colony forming ability and growth. Furthermore, treatment of fully formed, 6-day Prostaspheres for 48 hr with c-myc anti-sense significantly reduced c-myc expression and their ability to generate lesions in NOD-SCIDs (n = 10 Prostaspheres injected i.t./mouse).

Conclusions: These data demonstrate for the first time that a single CD133(hi) cell is competent to generate Prostaspheres in vitro and that CD133(hi) Prostaspheres require c-myc to grow and form dysplastic lesions in vivo.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • AC133 Antigen
  • Animals
  • Antigens, CD / biosynthesis*
  • Blotting, Western
  • DNA, Antisense / administration & dosage
  • DNA, Antisense / genetics*
  • Flow Cytometry
  • Glycoproteins / biosynthesis*
  • Humans
  • Karyotyping
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Middle Aged
  • Neoplastic Stem Cells / pathology*
  • Peptides
  • Prostatic Neoplasms / genetics
  • Prostatic Neoplasms / immunology
  • Prostatic Neoplasms / pathology*
  • Proto-Oncogene Proteins c-myc / biosynthesis
  • Proto-Oncogene Proteins c-myc / genetics*
  • Telomerase / genetics
  • Transfection

Substances

  • AC133 Antigen
  • Antigens, CD
  • DNA, Antisense
  • Glycoproteins
  • MYC protein, human
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse
  • Proto-Oncogene Proteins c-myc
  • TERT protein, human
  • Telomerase