Premium
Detection of cell‐free RNA in children with neuroblastoma and comparison with that of whole blood cell RNA
Author(s) -
Corrias Maria Valeria,
Pistorio Angela,
Cangemi Giuliana,
Tripodi Gino,
Carlini Barbara,
Scaruffi Paola,
Fardin Paolo,
Garaventa Alberto,
Pistoia Vito,
Haupt Riccardo
Publication year - 2010
Publication title -
pediatric blood and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.116
H-Index - 105
eISSN - 1545-5017
pISSN - 1545-5009
DOI - 10.1002/pbc.22498
Subject(s) - rna , cell , neuroblastoma , medicine , microbiology and biotechnology , gene , biology , cell culture , biochemistry , genetics
Background Since there is no validated assay to monitor disease in children with neuroblastoma (NB), we tested whether NB specific cell‐free RNA could be detected in their plasma samples. Moreover, with the aim of reducing patients' discomfort, we compared this assay to a recently standardized procedure that uses a larger amount of whole blood. Procedures Using conditions that excluded RNA recovery from contaminating tumor cells, the total amount of cell‐free RNA present in healthy children and patients with NB was quantified. Expression of tyrosine hydroxylase ( TH ) was assayed by quantitative RT‐PCR. Results In patients with NB the amount of cell‐free RNA was higher than in healthy children. However, it was less and more degraded than in healthy adults. The median amount of cell‐free RNA that was reverse transcribed, measured through the use of standard curves for reference genes, was 0.03 (range 0–30) pg of input RNA, that is, always less than 1/10,000 of that reverse transcribed from total RNA extracted from whole cells. Despite the presence of disease and the positive results obtained with RNA extracted from peripheral blood cells, few cell‐free RNA samples tested positive by the TH assay. Similar results were obtained also with TH primers specifically designed to amplify 50 bp RNA fragments. Conclusion These findings suggest that for monitoring disease status detection of cell‐free tumor‐specific RNAs in patients with NB is not a reliable alternative to whole cell RNA. Pediatr Blood Cancer 2010;54:897–903 © 2010 Wiley‐Liss, Inc.