Diegmann Julia, Junker Kerstin, Gerstmayer Bernhard, Bosio Andreas, Hindermann Winfried, Rosenhahn Julia, von Eggeling Ferdinand
Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, 07740 Jena, Germany.
Eur J Cancer. 2005 Aug;41(12):1794-801. doi: 10.1016/j.ejca.2005.05.005.
The underlying molecular mechanisms of renal cell carcinoma (RCC) are poorly understood and more reliable markers for early diagnosis are needed. Hence, alternative strategies for biomarker discovery with appropriate validation technologies have to be performed. To elucidate genesis and progression of RCC we used high parallel chip based gene expression profiling comparing normal and tumour tissues. We compared corresponding control and tumour tissue samples from 10 patients with clear cell RCC. We isolated RNA from histologically well characterised tissue sections and performed reverse transcription, labelling and linear RNA amplification. Samples were hybridised on microarrays containing 642 human cDNAs. Of the 352 differentially expressed genes found, CD70 and FRA2 were selected for further evaluation by real-time RT-PCR. The analysis all showed a high potential to discriminate between normal and tumour tissue. Moreover, increased CD70 mRNA expression in tumour cells could be correlated to its expression at the protein level. Immunohistochemistry (IHC) showed very strong expression of CD70 in all tumour samples but no expression in adjacent normal kidney tissue. With our combined approach we were able to identify CD70 as a new marker for RCC, which may be useful in the future for improved immunohistochemical diagnosis.
肾细胞癌（RCC）的潜在分子机制尚不清楚，因此需要更可靠的早期诊断标志物。因此，必须采用具有适当验证技术的生物标志物发现替代策略。为了阐明RCC的发生和发展，我们使用基于芯片的高通量基因表达谱分析来比较正常组织和肿瘤组织。我们比较了10例透明细胞RCC患者的相应对照组织和肿瘤组织样本。我们从组织学特征明确的组织切片中分离RNA，并进行逆转录、标记和线性RNA扩增。样本与包含642个人类cDNA的微阵列杂交。在发现的352个差异表达基因中，选择CD70和FRA2通过实时RT-PCR进行进一步评估。分析均显示出区分正常组织和肿瘤组织的高潜力。此外，肿瘤细胞中CD70 mRNA表达的增加与其蛋白质水平的表达相关。免疫组织化学（IHC）显示CD70在所有肿瘤样本中均有非常强的表达，但在相邻正常肾组织中无表达。通过我们的联合方法，我们能够将CD70鉴定为RCC的一种新标志物，这在未来可能有助于改进免疫组织化学诊断。
