Bjerregaard Henriette, Pedersen Shona, Kristensen Søren Risom, Marcussen Niels
Department of Clinical Biochemistry, Aalborg Hospital, Aarhus University Hospital, Aalborg, Denmark.
Diagn Mol Pathol. 2011 Dec;20(4):212-7. doi: 10.1097/PDM.0b013e318212e0a9.
Differentiation between malignant renal cell carcinoma and benign oncocytoma is of great importance to choose the optimal treatment. Accurate preoperative diagnosis of renal tumor is therefore crucial; however, existing imaging techniques and histologic examinations are incapable of providing an optimal differentiation profile. Analysis of gene expression of molecular markers is a new possibility but relies on appropriate standardization to compare different samples. The aim of this study was to identify stably expressed reference genes suitable for the normalization of results extracted from gene expression analysis of renal tumors.
Expression levels of 8 potential reference genes (ATP5J, HMBS, HPRT1, PPIA, TBP, 18S, GAPDH, and POLR2A) were examined by real-time reverse transcription polymerase chain reaction in tumor and normal tissue from removed kidneys from 13 patients with renal cell carcinoma and 5 patients with oncocytoma.
The expression levels of genes were compared by gene stability value M, average gene stability M, pairwise variation V, and coefficient of variation CV. More candidates were not suitable for the purpose, but a combination of HMBS, PPIA, ATP5J, and TBP was found to be the best combination with an average gene stability value M of 0.9 and a CV of 0.4 in the 18 tumors and normal tissues.
A combination of 4 genes, HMBS, PPIA, ATP5J, and TBP, is a possible reference in renal tumor gene expression analysis by reverse transcription polymerase chain reaction. A combination of four genes, HMBS, PPIA, ATP5J and TBP, being stably expressed in tissues from RCC is possible reference genes for gene expression analysis.
区分恶性肾细胞癌和良性嗜酸细胞瘤对于选择最佳治疗方案至关重要。因此，准确的肾肿瘤术前诊断至关重要；然而，现有的成像技术和组织学检查无法提供最佳的鉴别特征。分析分子标志物的基因表达是一种新的可能性，但依赖于适当的标准化来比较不同的样本。本研究的目的是确定适合对肾肿瘤基因表达分析结果进行标准化的稳定表达的参考基因。
通过实时逆转录聚合酶链反应检测了13例肾细胞癌患者和5例嗜酸细胞瘤患者切除肾脏的肿瘤组织和正常组织中8个潜在参考基因（ATP5J、HMBS、HPRT1、PPIA、TBP、18S、GAPDH和POLR2A）的表达水平。
通过基因稳定性值M、平均基因稳定性M、成对变异V和变异系数CV比较基因表达水平。更多的候选基因不适合该目的，但发现HMBS、PPIA、ATP5J和TBP的组合是最佳组合，在18个肿瘤组织和正常组织中平均基因稳定性值M为0.9，变异系数CV为0.4。
HMBS、PPIA、ATP5J和TBP这4个基因的组合可能是肾肿瘤逆转录聚合酶链反应基因表达分析的参考。HMBS、PPIA、ATP5J和TBP这4个基因的组合在肾细胞癌组织中稳定表达，可能是基因表达分析的参考基因。
