Molecular detection of neuron-specific ELAV-like-positive cells in the peripheral blood of patients with small-cell lung cancer.
Molecular detection of neuron-specific ELAV-like-positive cells in the peripheral blood of patients with small-cell lung cancer.
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DOI：
10.3233/clo-2008-0424
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Vendemiale G
中科院分区：
文献类型：
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作者：
D'Alessandro V;Muscarella LA;Copetti M;Zelante L;Carella M;Vendemiale G
背景：n-ELAV（neuronal-Embryonic Lethal，Abnormal Vision）样基因是编码肿瘤RNA结合蛋白的家族。抗-Hu-抗体（抗-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）中发现低滴度的抗-Hu-Ab。本研究的目的是建立一种灵敏、定量的分子实时定量PCR方法，通过定量nELAV样转录本检测外周血中的SCLC细胞。 方法：采用实时荧光PCR技术检测25例SCLC患者和12例健康献血员外周血标本。使用质粒稀释液作为校准曲线，用绝对定量方法测量外周血样品中HuB（ELAV 2）、HuC（ELAV 3）和HuD（ELAV 4）的mRNA水平。 结果如下：与来自健康献血者的样品相比，在SCLC患者中检测到HuB和HuD的mRNA表达水平的统计学显著增加。在基于对照样品中的表达水平建立截止值后，28%的SCLC样品对HuD表达呈阳性。总体而言，60%的SCLC显示HuD或HuB转录物水平增加。 结论：我们的初步结果表明，神经元-ELAV mRNA检测SCLC患者外周血中使用实时定量PCR。
Background: 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. Methods: Peripheral blood samples from 25 SCLC untreated patients and 12 healthy blood donors were investigated by realtime 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. Results: 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. Conclusion: Our preliminary results suggest that neuron-ELAV mRNA are detectable in peripheral blood of SCLC patients using real-time quantitative PCR.