D'Alessandro Vito, Muscarella Lucia Anna, Copetti Massimiliano, Zelante Leopoldo, Carella Massimo, Vendemiale Gianluigi
Internal Medicine Department, IRCCS Casa Sollievo della Sofferenza Hospital, San Giovanni Rotondo (FG), Italy.
Cell Oncol. 2008;30(4):291-7. doi: 10.3233/clo-2008-0424.
n-ELAV (neuronal-Embryonic Lethal, Abnormal Vision)-like genes belong to a family codifying for onconeural RNA-binding proteins. Anti-Hu-antibodies (anti-Hu-Ab) are typically associated with paraneoplastic encephalomyelitis/sensory neuropathy (PEM/PSN), and low titres of anti-Hu-Ab, were found in newly diagnosed Small Cell Lung Cancer (SCLC). The aim of this study is to develop a sensitive and quantitative molecular real-time PCR assay to detect SCLC cells in peripheral blood (PB) through nELAV-like transcripts quantification.
Peripheral blood samples from 25 SCLC untreated patients and 12 healthy blood donors were investigated by real-time PCR. mRNA levels for HuB (ELAV2), HuC (ELAV3) and HuD (ELAV4) were measured in peripheral blood samples with an absolute quantification method using plasmid dilutions as calibration curves.
A statistically significant increase in mRNA expression level was detected for HuB and HuD in SCLC patients as compared with samples from healthy blood donors. After establishing cut off values based on the level of expression in control samples, 28% of the SCLC samples were positive for HuD expression. Overall 60% of the SCLC displayed increased level of HuD or HuB transcripts.
Our preliminary results suggest that neuron-ELAV mRNA are detectable in peripheral blood of SCLC patients using real-time quantitative PCR.
神经元胚胎致死性异常视觉(n-ELAV)样基因属于一个编码癌胚RNA结合蛋白的家族。抗Hu抗体(anti-Hu-Ab)通常与副肿瘤性脑脊髓炎/感觉神经病变(PEM/PSN)相关,并且在新诊断的小细胞肺癌(SCLC)中发现了低滴度的抗Hu-Ab。本研究的目的是开发一种灵敏且定量的分子实时PCR检测方法,通过定量nELAV样转录本来检测外周血(PB)中的SCLC细胞。
采用实时PCR对25例未经治疗的SCLC患者和12名健康献血者的外周血样本进行研究。使用质粒稀释作为标准曲线的绝对定量方法,测量外周血样本中HuB(ELAV2)、HuC(ELAV3)和HuD(ELAV4)的mRNA水平。
与健康献血者的样本相比,SCLC患者中HuB和HuD的mRNA表达水平有统计学意义的升高。根据对照样本的表达水平确定临界值后,28%的SCLC样本HuD表达呈阳性。总体而言,60%的SCLC显示HuD或HuB转录本水平升高。
我们的初步结果表明,使用实时定量PCR可在SCLC患者外周血中检测到神经元ELAV mRNA。