Kunth K, Höfler H, Atkinson M J
GSF-Forschungzentrum für Umwelt und Gesundheit, Institut für Pathologie, Oberschlessheim.
Verh Dtsch Ges Pathol. 1994;78:226-30.
Alterations in gene expression are frequently encountered in malignant tissues. Quantification of the often quite subtle changes in mRNA content is routinely performed by northern blot hybridization. Practical experience shows that the methodology is subject to considerable error, necessitating the use of an internal mRNA standard for quantification. Many different control mRNAs have been employed over the years, but no systematic study has proven their applicability to comparisons made between tumor cells and normal tissues. The validity of these control RNAs, usually housekeeping genes, must be established to exclude the possibility that they themselves are expressed at different levels in tumor and non-tumor cells. Our experience has revealed that serious discrepancies may arise when incorrect standard genes are used to compare tumor and normal tissues.
基因表达的改变在恶性组织中经常出现。通过Northern印迹杂交可常规检测mRNA含量中通常相当细微的变化。实际经验表明，该方法存在相当大的误差，因此需要使用内部mRNA标准进行定量。多年来已采用了许多不同的对照mRNA，但尚无系统研究证明它们适用于肿瘤细胞与正常组织之间的比较。必须确定这些对照RNA（通常是管家基因）的有效性，以排除它们自身在肿瘤细胞和非肿瘤细胞中表达水平不同的可能性。我们的经验表明，使用错误的标准基因来比较肿瘤组织和正常组织时可能会出现严重差异。
