Kuroda T, Saeki M, Nakano M, Mizutani S
Department of Surgery, National Children's Hospital, Tokyo, Japan.
J Pediatr Surg. 1997 Jan;32(1):69-72. doi: 10.1016/s0022-3468(97)90097-x.
The highly sensitive method to detect neuroblastoma (NB) cells using reverse transcriptase polymerase chain reaction (RT-PCR) was applied in the practical clinics, and its efficacy was assessed in the present study. Human tyrosine hydroxylase (TH), a rate-limiting enzyme in the catecholamine biosynthesis, was used as the marker for NB cells, and the expression of THmRNA was examined in 13 samples (four peripheral blood and nine bone marrow) harvested from seven patients (four with stage IV, one with stage III, two with stage II) using RT-PCR with our original primers. The positive signals for NB cells were detected in four samples (one peripheral blood and three bone marrow) by the PCR method, but were undetectable by the conventional histological examinations. In the present series, a case that showed a positive signal for NB cells in the peripheral blood showed a remarkably unfavorable clinical course, indicating that the circulating NB cells detected by the PCR method can be a sign of the progressively advanced NB, and may define a new prognostic factor suggesting higher risk. In another case, the PCR detection for the residual NB cells in the bone marrow provided important supporting evidence to determine the necessity of the additional chemotherapy and the suitable timing for bone marrow transplantation. This detection also guaranteed the safety of the bone marrow for transplantation. The PCR method was considered to be very beneficial in the selected cases. However, some problems such as the false-negative results in the negative urinary vanillylmandelic acid secretor were also highlighted in the present study.
本研究应用逆转录聚合酶链反应（RT-PCR）检测神经母细胞瘤（NB）细胞的高灵敏度方法，并在实际临床中进行了疗效评估。人酪氨酸羟化酶（TH）是儿茶酚胺生物合成中的限速酶，用作NB细胞的标志物，使用我们的原始引物通过RT-PCR检测了从7例患者（4例IV期、1例III期、2例II期）采集的13份样本（4份外周血和9份骨髓）中THmRNA的表达。通过PCR方法在4份样本（1份外周血和3份骨髓）中检测到NB细胞的阳性信号，但常规组织学检查未检测到。在本系列中，1例在外周血中显示NB细胞阳性信号的病例临床病程明显不佳，表明通过PCR方法检测到的循环NB细胞可能是NB进展的标志，可能定义一个提示更高风险的新预后因素。在另一例中，对骨髓中残留NB细胞的PCR检测为确定是否需要额外化疗以及骨髓移植的合适时机提供了重要的支持证据。这种检测还确保了用于移植的骨髓的安全性。PCR方法被认为在选定病例中非常有益。然而，本研究也突出了一些问题，如尿香草扁桃酸分泌阴性者出现假阴性结果。
