黑色素瘤中基因表达数据的标准化：研究将3-磷酸甘油醛脱氢酶和18S核糖体RNA用作定量实时PCR的内参基因
The normalization of gene expression data in melanoma: investigating the use of glyceraldehyde 3-phosphate dehydrogenase and 18S ribosomal RNA as internal reference genes for quantitative real-time PCR.
作者信息
Giricz Orsolya, Lauer-Fields Janelle L, Fields Gregg B
机构信息
Department of Chemistry and Biochemistry, Florida Atlantic University, 777 Glades Road, Boca Raton, FL 33431-0991, USA.
出版信息
Anal Biochem. 2008 Sep 1;380(1):137-9. doi: 10.1016/j.ab.2008.05.024. Epub 2008 May 22.
Abstract
We examined a panel of 26 melanoma and fibroblast samples (tissues and cultured cells) to evaluate the suitability of two commonly used housekeeping genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and 18S ribosomal RNA (rRNA), for quantitative real-time PCR. Both genes showed significant variations within the individual cell line and tissue groups. Although no overall trends were observed in the expression of the 18S rRNA, GAPDH was up-regulated in melanoma tissue and cultured cells compared with the corresponding normal samples. In melanoma and fibroblast cell lines and tissues, absolute quantification appears to be more appropriate than normalizing messenger RNA (mRNA) expression via GAPDH or 18S rRNA housekeeping genes.
摘要
我们检测了一组26个黑色素瘤和成纤维细胞样本（组织和培养细胞），以评估两种常用管家基因——甘油醛-3-磷酸脱氢酶（GAPDH）和18S核糖体RNA（rRNA）用于定量实时PCR的适用性。这两个基因在各个细胞系和组织组中均表现出显著差异。虽然在18S rRNA的表达中未观察到总体趋势，但与相应的正常样本相比，GAPDH在黑色素瘤组织和培养细胞中上调。在黑色素瘤和成纤维细胞系及组织中，绝对定量似乎比通过GAPDH或18S rRNA管家基因对信使RNA（mRNA）表达进行标准化更为合适。
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