Friedrich M G, Erbersdobler A, Schwaibold H, Conrad S, Huland E, Huland H
Department of Urology and Institute for Pathology, University Hospital Eppendorf, University of Hamburg, Germany.
J Urol. 2000 Mar;163(3):1039-42.
Detection of loss of heterozygosity (LOH) has been described in various carcinomas on the basis of meticulous molecular techniques. Because of lack of simple and rapid techniques, LOH has not achieved common use in routine tumor diagnosis. A recently found variable number of tandem repeats (VNTR) segment in intron 1 of the p53 gene was described as highly polymorphic and therefore useful in detecting LOH. We used a rapid technique for detection of LOH in the p53 gene of patients with transitional cell carcinoma (TCC) of the bladder. The technique was based on the polymerase chain reaction (PCR) and agarose gel electrophoresis as described for other carcinomas previously. We evaluated whether TCC screening and surveillance could be performed detecting LOH in the urinary sediment.
We investigated 29 patients with TCC of the bladder (pTa 12 patients; pT1 10 patients; pT2 - pT4 seven patients; grade 1 one patient; grade 2 19 patients; grade 3 nine patients). DNA was prepared by standard methods from white blood cells, tumor tissue, normal bladder mucosa, and urinary sediments. The amplification of the VNTR region was performed with PCR. PCR products were run in parallel lanes on 4.5% agarose gels.
Of the 29 patients, 23 (79.3%) were found to have two different alleles ("informative cases") for the VNTR region. Of the 23 informative cases LOH was detected in the tumor tissue of 10 patients (43.5%). Referring to the total population 10 of 29 patients (34.4%) revealed LOH. In all patients with LOH in the tumor, LOH was also detected in the urinary sediment. LOH was not detected in the histologically benign bladder mucosa.
We present a simple and rapid technique based on PCR and agarose gel electrophoresis for the detection of LOH in tumor and urinary sediment of patients with TCC of the bladder. The ability to detect LOH not only in tumor tissue but also in urinary sediment offers an attractive approach for noninvasive diagnosis and surveillance of bladder cancer patients.
基于精细的分子技术，已经在多种癌症中描述了杂合性缺失（LOH）的检测。由于缺乏简单快速的技术，LOH尚未在常规肿瘤诊断中得到广泛应用。最近在p53基因内含子1中发现的可变串联重复序列（VNTR）片段被描述为高度多态性，因此可用于检测LOH。我们采用一种快速技术检测膀胱移行细胞癌（TCC）患者p53基因中的LOH。该技术基于聚合酶链反应（PCR）和琼脂糖凝胶电泳，如先前针对其他癌症所描述的那样。我们评估了是否可以通过检测尿沉渣中的LOH来进行TCC筛查和监测。
我们研究了29例膀胱TCC患者（pTa期12例；pT1期10例；pT2 - pT4期7例；1级1例；2级19例；3级9例）。通过标准方法从白细胞、肿瘤组织、正常膀胱黏膜和尿沉渣中提取DNA。使用PCR对VNTR区域进行扩增。PCR产物在4.5%琼脂糖凝胶的平行泳道中进行电泳。
在29例患者中，23例（79.3%）的VNTR区域存在两种不同的等位基因（“信息性病例”）。在这23例信息性病例中，10例患者（43.5%）的肿瘤组织中检测到LOH。在全部29例患者中，10例（34.4%）存在LOH。在所有肿瘤中存在LOH的患者的尿沉渣中也检测到了LOH。在组织学上良性的膀胱黏膜中未检测到LOH。
我们提出了一种基于PCR和琼脂糖凝胶电泳的简单快速技术，用于检测膀胱TCC患者肿瘤组织和尿沉渣中的LOH。不仅能够在肿瘤组织中，而且能够在尿沉渣中检测到LOH，为膀胱癌患者的无创诊断和监测提供了一种有吸引力的方法。
