Etschmann B, Wilcken B, Stoevesand K, von der Schulenburg A, Sterner-Kock A
Institut für Veterinaerphysiologie, Fachbereich Veterinaermedizin, Freie Universitaet Berlin, Germany.
Vet Pathol. 2006 Nov;43(6):934-42. doi: 10.1354/vp.43-6-934.
Eleven reference genes (18s ribosomal ribonucleic acid [RNA], 28s ribosomal RNA, ubiquitin, beta-actin, glycerine aldehyde dehydrogenase, ATP-synthase subunit 5B, hydroxymethyl-bilane synthase, hypoxanthine-phosphoribosyl transferase, ribosomal protein L32, tryptophan 5-monooxygenase activation protein (zeta polypeptide), and TATA-Box binding protein) were analyzed in use as references for gene expression profiling experiments using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in canine mammary tumors. The transcription level of the candidates was measured in 22 histologically characterized excised tumor specimens from mammary gland tissue and 22 samples of non-neoplastic mammary tissue samples from the same individuals. Results were used to rank candidate reference genes using the GeNorm tool. It was determined that in samples of canine mammary gland tissue, a combination of hypoxanthine-phosphoribosyl transferase, ATP-synthase subunit 5B, ribosomal protein L32 and ubiquitin yields stable reference gene expression levels, whereas the use of glycerin aldehyde dehydrogenase or ribosomal RNA is unsuitable for normalization of qRT-PCR results in this tissue type.
在犬乳腺肿瘤中，对11个参考基因（18s核糖体核糖核酸[RNA]、28s核糖体RNA、泛素、β-肌动蛋白、甘油醛脱氢酶、ATP合酶亚基5B、羟甲基胆色素合酶、次黄嘌呤-磷酸核糖基转移酶、核糖体蛋白L32、色氨酸5-单加氧酶激活蛋白（ζ多肽）和TATA盒结合蛋白）进行了分析，以用作使用定量逆转录聚合酶链反应（qRT-PCR）进行基因表达谱实验的参考。在22个来自乳腺组织的经组织学鉴定的切除肿瘤标本和来自同一动物个体的22个非肿瘤性乳腺组织样本中测量了候选基因的转录水平。使用GeNorm工具对候选参考基因进行排名。结果表明，在犬乳腺组织样本中，次黄嘌呤-磷酸核糖基转移酶、ATP合酶亚基5B、核糖体蛋白L32和泛素的组合产生稳定的参考基因表达水平，而甘油醛脱氢酶或核糖体RNA不适用于该组织类型的qRT-PCR结果的标准化。
