Dysplasia of human prostate CD133(hi) sub-population in NOD-SCIDS is blocked by c-myc anti-sense.
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.
Goodyear
S M
SM
Molecular Pathobiology Program, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA.
Amatangelo
M D
MD
Stearns
M E
ME
D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D015153:Blotting, Western; D016373:DNA, Antisense / Q000008:administration & dosage / Q000235:genetics*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000096:biosynthesis*; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000473:pathology*; D010455:Peptides; D011471:Prostatic Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000235:genetics*; D019098:Telomerase / Q000235:genetics; D014162:Transfection
D000071916:AC133 Antigen; D015703:Antigens, CD; D016373:DNA, Antisense; D006023:Glycoproteins; C489427:MYC protein, human; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; C509186:TERT protein, human; D019098:Telomerase