A distinct gene expression signature characterizes human neuroblastoma cancer stem cells.
Neuroblastoma, a malignancy of multipotent embryonic neural crest cells, is the most common extracranial solid cancer in childhood and most common cancer in infancy. Cellular phenotype has been shown to be an important determinant of the malignant potential in human neuroblastoma cells and tumors. Whereas neuroblastic (N-type) are moderately malignant and nonneuronal (S-type) cells are nonmalignant, I-type stem cells are highly tumorigenic, irrespective of N-myc amplification status. In the present study, we sought to determine which genes were overexpressed in the I-type cells which might characterize and maintain the stem cell state and/or malignancy of human neuroblastoma cancer stem cells. We used a microarray platform to compare the steady-state expression levels of mRNAs from 13 human neuroblastoma cell lines representing the three cellular phenotypes. Using qRT-PCR and Western blot analyses, we identified seven genes whose expression is consistently elevated exclusively in neuroblastoma cancer stem cells: CD133, KIT, NOTCH1, GPRC5C, PIGF2, TRKB, and LNGFR. Moreover, we show that the genes are phenotype specific, as differentiation of I-type BE(2)-C cells to either an N- or S-type morphology results in significantly reduced mRNA expression. Finally, we show that NOTCH1 plays an important role in maintaining the stem cell phenotype. The identification and characterization of these genes, elevated in highly malignant neuroblastoma stem cells, could provide the basis for developing novel therapies for treatment of this lethal childhood cancer.
Laboratory of Neurobiology, Department of Biological Sciences, Fordham University, 441 E. Fordham Rd., Bronx, NY 10458, United States. Electronic address: rross@fordham.edu.
Robert A
RA
Ross
Laboratory of Neurobiology, Department of Biological Sciences, Fordham University, 441 E. Fordham Rd., Bronx, NY 10458, United States; Department of Biological Sciences, Seton Hall University, 400 South Orange Ave., South Orange, NJ 07079, United States.
Jeanette D
JD
Walton
Laboratory of Neurobiology, Department of Biological Sciences, Fordham University, 441 E. Fordham Rd., Bronx, NY 10458, United States.
Dan
D
Han
Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, United States.
Hong-Fen
HF
Guo
Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, United States.
Nai-Kong V
NK
Cheung
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000378:metabolism; D010641:Phenotype; D000072483:Placenta Growth Factor; D011257:Pregnancy Proteins / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; C417919:GPRC5C protein, human; D006023:Glycoproteins; C000608036:PGF protein, human; C000606023:PROM1 protein, human; D010455:Peptides; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051881:Receptor, Notch1; D043562:Receptors, G-Protein-Coupled; D000072483:Placenta Growth Factor; D019009:Proto-Oncogene Proteins c-kit
Differentiation; GPRC5C; Neuroblastoma; Neurotrophic receptors; Notch1; Placental growth factor-2; Stem cell