Detection of neuroblastoma cells in blood by reverse transcriptase-polymerase chain reaction.
OBJECTIVE
To determine whether circulating tumor cells are present in patients with localized or disseminated neuroblastoma (NB).


METHODS
Sixteen pediatric malignant tumor cell lines, including 10 NB, 4 primitive neuroectodermal tumor (PENT), 2 Wilms' tumor and 39 samples (tumor tissue, bone marrow and peripheral blood) from 8 patients with NB, were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) using two markers: neuroendocrine protein gene products 9.5 (PGP 9.5) and tyrosine hydroxylase (TH).


RESULTS
Analysis of RT-PCR products by agarose electrophoresis demonstrated that in culture cell lines the specificity for NB was highest in TH (9/10 with NB, 3/4 with PNET, 0/2 with Wilms' tumor) and the sensitivity was highest in PGP 9.5 10(-7). PGP 9.5 and TH mRNA were undetectable in the peripheral blood mononuclear cells (PBMNC) from normal children. Expression of these markers in 8 patients with NB at diagnosis and during treatment was found positive in 5 out of 8, all had been proved paralleling with their disease status. Two of the 5 became negative after further treatment.


CONCLUSIONS
These systems allow the detection of circulating NB cells with higher sensitivity and specificity. When RT-PCR is clinically used for the detection of NB cells in blood, that both PGP 9.5 and TH should be selected is recommended. The assessment of bone marrow alone might not be used as a single index of tumor dissemination.
Gao
Y
Y
Department of Pediatric Surgery, Second Affiliated Hospital of Xi'an Medical University, China.
Li
G
G
Zhang
X
X
Xu
Q
Q
Zheng
B
B
D014408:Biomarkers, Tumor / Q000032:analysis; D001932:Brain Neoplasms / Q000473:pathology*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D007680:Kidney Neoplasms / Q000473:pathology; D008297:Male; D009360:Neoplastic Cells, Circulating; D009419:Nerve Tissue Proteins / Q000032:analysis; D009447:Neuroblastoma / Q000473:pathology*; D018242:Neuroectodermal Tumors, Primitive / Q000473:pathology*; D016133:Polymerase Chain Reaction / Q000379:methods; D013869:Thiolester Hydrolases / Q000032:analysis; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis; D043222:Ubiquitin Thiolesterase; D009396:Wilms Tumor / Q000473:pathology
D014408:Biomarkers, Tumor; D009419:Nerve Tissue Proteins; D014446:Tyrosine 3-Monooxygenase; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase