Schmidt Bernd, Engel Eileen, Carstensen Tim, Weickmann Sabine, John Matthias, Witt Christian, Fleischhacker Michael
Division of Pneumology, University Hospital Charité, Charité - Universitätsmedizin Berlin, Campus Charité-Mitte, Schumannstr. 20-21, D-10117 Berlin, Germany.
Lung Cancer. 2005 Apr;48(1):145-7. doi: 10.1016/j.lungcan.2004.09.013. Epub 2004 Dec 16.
Circulating cell-free nucleic acids have been detected in serum. In cancer patients levels of free DNA seem to be higher than in non-tumor controls and the detectable nucleic acids are partly of tumor origin. We asked whether free RNA can be detected as well in cell-free bronchial lavage fluid (BLF) supernatant, and whether quantification of free RNA allows to discriminate between tumor and non-tumor patients. 73 patients with lung cancer (NSCLC n=62, SCLC n=11) and 56 patients with non-malignant lung diseases were included. The RNA was isolated from 1 mL serum and lavage, respectively with the Quiamp MinElute Virus Vacuum Kit (Qiagen, Germany). A real-time quantitative RT-PCR assay was used for transcript quantification of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene. Intact RNA was detectable in cell-free supernatants of BLF from 126/129 patients and was investigated in all 64 serum samples. RNA levels were higher in the cell-free supernatant of BLF than in serum. RNA concentration in the BLF from tumor patients was higher than in patients with a benign lung disease (p=0.009). In conclusion, quantification of intact RNA isolated from BLF supernatant and from serum might become a valuable tool for differentiation between tumor and non-tumor patients.
血清中已检测到循环游离核酸。在癌症患者中,游离DNA水平似乎高于非肿瘤对照,且可检测到的核酸部分来源于肿瘤。我们询问在游离支气管灌洗液(BLF)上清液中是否也能检测到游离RNA,以及游离RNA的定量分析是否有助于区分肿瘤患者和非肿瘤患者。纳入了73例肺癌患者(非小细胞肺癌62例,小细胞肺癌11例)和56例非恶性肺部疾病患者。分别使用Quiamp MinElute病毒真空试剂盒(德国Qiagen公司)从1 mL血清和灌洗液中分离RNA。采用实时定量逆转录聚合酶链反应(RT-PCR)法对甘油醛-3-磷酸脱氢酶(GAPDH)基因进行转录本定量分析。129例患者中有126例的BLF游离上清液中可检测到完整RNA,并对所有64份血清样本进行了研究。BLF游离上清液中的RNA水平高于血清。肿瘤患者BLF中的RNA浓度高于良性肺部疾病患者(p = 0.009)。总之,对从BLF上清液和血清中分离出的完整RNA进行定量分析可能成为区分肿瘤患者和非肿瘤患者的有价值工具。