McNulty S E, Toscano W A
Center for Bioenvironmental Research, Tulane University School of Public Health and Tropical Medicine, New Orleans, LA 70112-2699, USA.
Biochem Biophys Res Commun. 1995 Jul 6;212(1):165-71. doi: 10.1006/bbrc.1995.1951.
The action of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) on transcription and steady-state mRNA levels, protein expression, and enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cultured human keratinocytes was examined. GAPDH transcription increased as a function of time of exposure to TCDD, reaching a maximum of 23-fold after 8 days. Northern blot analysis showed a 4-fold increase in mRNA after 8 days exposure. Protein levels were increased 4-fold, and dehydrogenase activity by 5-fold, under the same exposure conditions. GAPDH is commonly thought of as a constitutive housekeeping gene and is often used as a loading control for northern blot analysis. Our data suggest that caution should be used when using GAPDH as an RNA control, especially in studies examining cell proliferation and carcinogenesis.
研究了2,3,7,8-四氯二苯并-对-二恶英(TCDD)对培养的人角质形成细胞中甘油醛-3-磷酸脱氢酶(GAPDH)的转录、稳态mRNA水平、蛋白质表达和酶活性的作用。GAPDH转录随TCDD暴露时间的增加而增加,8天后达到最大23倍。Northern印迹分析显示,暴露8天后mRNA增加了4倍。在相同的暴露条件下,蛋白质水平增加了4倍,脱氢酶活性增加了5倍。GAPDH通常被认为是一个组成型管家基因,常用于Northern印迹分析的上样对照。我们的数据表明,在将GAPDH用作RNA对照时应谨慎,尤其是在研究细胞增殖和致癌作用的研究中。