In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR.
In search of suitable reference genes for gene expression studies of human renal cell carcinoma by real-time PCR.
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DOI：
10.1186/1471-2199-8-47
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发表时间：
2007-06-08
影响因子：
--
通讯作者：
Jung K
中科院分区：
文献类型：
--
作者：
Jung M;Ramankulov A;Roigas J;Johannsen M;Ringsdorf M;Kristiansen G;Jung K
管家基因通常用作基因表达研究中目标基因相对定量的内源参考基因。迄今为止，尚未发表比较不同候选参考基因在肾透明细胞癌中的适用性的结论性系统研究。为了补救这种情况，10管家基因的RT-PCR测量已经在各种研究中推荐的标准化的目的进行了检查，就其作为参考基因的有用性。在25例肾透明细胞癌患者的恶性和非恶性组织标本中检测潜在参考基因的表达。用2100 Agilent Bioanalyzer进行的分离RNA的质量评估显示所有样品的平均RNA完整性数值为8.7。与对照材料的PCR反应交叉点相关的运行间变异范围为0.17%至0.38%。所有基因的表达不依赖于年龄、性别和肿瘤分期。除TATA盒结合蛋白（TBP）和肽基脯氨酰异构酶A（PPIA）基因外，所有基因在恶性和非恶性配对中的表达均存在显著差异。另外使用软件程序geNorm和Normalware控制候选参考基因的表达稳定性。通过使用这两种最稳定表达的基因与该组的上调和下调的管家基因相比归一化靶基因ADAM 9，验证TBP和PPIA作为合适的参考基因。我们的研究表明，两个管家基因PPIA和TBP作为内源性参考基因的适用性时，比较恶性组织样本与相邻的正常组织样本从透明细胞肾细胞癌。这两个基因都被推荐作为参考基因，用于基因谱研究中的相对基因定量，无论是作为单一基因还是优选组合。
Housekeeping genes are commonly used as endogenous reference genes for the relative quantification of target genes in gene expression studies. No conclusive systematic study comparing the suitability of different candidate reference genes in clear cell renal cell carcinoma has been published to date. To remedy this situation, 10 housekeeping genes for normalizing purposes of RT-PCR measurements already recommended in various studies were examined with regard to their usefulness as reference genes. The expression of the potential reference genes was examined in matched malignant and non-malignant tissue specimens from 25 patients with clear cell renal cell carcinoma. Quality assessment of isolated RNA performed with a 2100 Agilent Bioanalyzer showed a mean RNA integrity number of 8.7 for all samples. The between-run variations related to the crossing points of PCR reactions of a control material ranged from 0.17% to 0.38%. The expression of all genes did not depend on age, sex, and tumour stage. Except the genes TATA box binding protein (TBP) and peptidylprolyl isomerase A (PPIA), all genes showed significant differences in expression between malignant and non-malignant pairs. The expression stability of the candidate reference genes was additionally controlled using the software programs geNorm and NormFinder. TBP and PPIA were validated as suitable reference genes by normalizing the target gene ADAM9 using these two most stably expressed genes in comparison with up- and down-regulated housekeeping genes of the panel. Our study demonstrated the suitability of the two housekeeping genes PPIA and TBP as endogenous reference genes when comparing malignant tissue samples with adjacent normal tissue samples from clear cell renal cell carcinoma. Both genes are recommended as reference genes for relative gene quantification in gene profiling studies either as single gene or preferably in combination.
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影响因子：
14.9
作者：
Imbeaud S;Graudens E;Boulanger V;Barlet X;Zaborski P;Eveno E;Mueller O;Schroeder A;Auffray C
通讯作者：
Auffray C
DOI：
10.2165/00066982-200408020-00005
发表时间：
2004-01-01
期刊：
Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology
影响因子：
--
作者：
Janssens, Nico;Janicot, Michel;Bakker, Annette
通讯作者：
Bakker, Annette
影响因子：
2.9
作者：
Haller, F;Kulle, B;Füzesi, L
通讯作者：
Füzesi, L
影响因子：
10
作者：
Glare, EM;Divjak, M;Walters, EH
通讯作者：
Walters, EH
影响因子：
3.5
作者：
Bustin, SA
通讯作者：
Bustin, SA