Köhler Christina U, Walter Michael, Lang Kerstin, Plöttner Sabine, Roghmann Florian, Noldus Joachim, Tannapfel Andrea, Tam Yu Chun, Käfferlein Heiko U, Brüning Thomas
Institute for Prevention and Occupational Medicine of the German Social Accident Insurance, Institute of the Ruhr University Bochum (IPA), Bürkle-de-la-Camp Platz 1, 44789 Bochum, Germany.
C.ATG Core Facility for NGS and Microarrays, University of Tübingen, Calwerstr. 7, 72076 Tübingen, Germany.
Biomedicines. 2020 Jul 22;8(8):233. doi: 10.3390/biomedicines8080233.
We identified DNA methylation targets specific for urothelial cancer (UC) by genome-wide methylation difference analysis of human urothelial (RT4, J82, 5637), prostate (LNCAP, DU-145, PC3) and renal (RCC-KP, CAKI-2, CAL-54) cancer cell lines with their respective primary epithelial cells. A large overlap of differentially methylated targets between all organs was observed and 40 Cytosine-phosphate-Guanine motifs (CpGs) were only specific for UC cells. Of those sites, two also showed high methylation differences (≥47%) in vivo when we further compared our data to those previously obtained in our array-based analyses of urine samples in 12 UC patients and 12 controls. Using mass spectrometry, we finally assessed seven CpG sites in this "bladder-specific" region of interest in urine samples of patients with urothelial ( = 293), prostate ( = 75) and renal ( = 23) cancer, and 143 controls. DNA methylation was significantly increased in UC compared to non-UC individuals. The differences were more pronounced for males rather than females. Male UC cases could be distinguished from non-UC individuals with >30% sensitivity at 95% specificity (Area under the curve (AUC) 0.85). In summary, methylation sites highly specific in UC cell lines were also specific in urine samples of UC patients showing that in-vitro data can be successfully used to identify biomarker candidates of in-vivo relevance.
通过对人尿路上皮（RT4、J82、5637）、前列腺（LNCAP、DU - 145、PC3）和肾（RCC - KP、CAKI - 2、CAL - 54）癌细胞系及其各自的原代上皮细胞进行全基因组甲基化差异分析，我们确定了尿路上皮癌（UC）特有的DNA甲基化靶点。观察到所有器官之间差异甲基化靶点有大量重叠，并且40个胞嘧啶 - 磷酸 - 鸟嘌呤基序（CpG）仅对UC细胞具有特异性。在这些位点中，当我们将我们的数据与之前在12名UC患者和12名对照的尿液样本基于阵列的分析中获得的数据进一步比较时，其中两个位点在体内也显示出高甲基化差异（≥47%）。我们最终使用质谱法评估了尿路上皮癌（n = 293）、前列腺癌（n = 75）和肾癌（n = 23）患者以及143名对照的尿液样本中这个“膀胱特异性”感兴趣区域的7个CpG位点。与非UC个体相比，UC患者的DNA甲基化显著增加。男性的差异比女性更明显。男性UC病例与非UC个体可以区分，在95%特异性时灵敏度>30%（曲线下面积（AUC）0.85）。总之，在UC细胞系中高度特异的甲基化位点在UC患者的尿液样本中也具有特异性，这表明体外数据可以成功用于识别具有体内相关性的生物标志物候选物。
