DETECTION OF NEUROBLASTOMA-CELLS IN BONE-MARROW AND PERIPHERAL-BLOOD AT DIAGNOSIS BY THE REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION FOR TYROSINE-HYDROXYLASE MESSENGER-RNA
Y. Miyajima et al., DETECTION OF NEUROBLASTOMA-CELLS IN BONE-MARROW AND PERIPHERAL-BLOOD AT DIAGNOSIS BY THE REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION FOR TYROSINE-HYDROXYLASE MESSENGER-RNA, Cancer, 75(11), 1995, pp. 2757-2761
Background, Bone marrow metastasis often occurs in patients with neuro
blastoma; therefore, a sensitive assay to detect occult neuroblastoma
cells in bone marrow (BM) and peripheral blood (PB) is needed, The fea
sibility and clinical value of using the reverse transcriptase-(RT) po
lymerase chain reaction (PCR) to amplify mRNA for tyrosine hydroxylase
(TH), the first enzyme of catecholamine synthesis, was evaluated to d
etect neuroblastoma cells in patient samples. Methods. Thirty-eight pa
tients with Stages I to IV neuroblastoma and eight healthy donors were
included in this study. Bone marrow and PB samples obtained at diagno
sis were examined for TH mRNA. After preparation of complementary DNA,
the PCR was performed to amplify the TH gene. Results. Tyrosine hydro
xylase mRNA was detected in neuroblastoma samples including a cell lin
e and tumor tissues, but was not detected in normal BM or PB mononucle
ar cells, Neuroblastoma cells were detected at a level of 1 per 10(5-6
) normal PB mononuclear cells by this method, Tyrosine hydroxylase mRN
A was detected in 18 of 38 BM samples, and all 12 BM samples with cyto
logic evidence of tumor cells were positive for TH mRNA by the RT-PCR,
Six of 26 patients without cytologic evidence of tumor cells in the B
M were also positive for TH mRNA. TH mRNA was detected in BM samples f
rom 1 of 14 patients with Stage I disease, 2 of 7 patients with Stage
II disease, 1 of 3 patients with Stage III disease and all patients wi
th Stage IV (11 patients) and IVS (3 patients) diseases. Tyrosine hydr
oxylase mRNA also was detected in 8 of 14 PB samples (one of five pati
ents in Stages I, II or III, and 7 of 9 in Stage IV or IVS). Conclusio
ns. Reverse transcriptase-polymerase chain reaction amplification of T
H mRNA was a sensitive and specific method of detecting occult neurobl
astoma cells in BM and PB samples, Neuroblastoma cells could be detect
ed by this method in some BM samples that had no cytologic evidence of
tumor cells and in some PB samples at the time of diagnosis. The clin
ical significance of these very low levels of neuroblastoma cells dete
cted by RT-PCR requires further investigation.