Jóźwik Maciej, Sidorkiewicz Iwona, Wojtkiewicz Joanna, Sulkowski Stanisław, Semczuk Andrzej, Jóźwik Marcin
Department of Gynecology and Gynecologic Oncology, Medical University of Białystok, 15-276 Białystok, Poland.
Clinical Research Support Centre, Medical University of Białystok, 15-276 Białystok, Poland.
Int J Mol Sci. 2025 Sep 4;26(17):8610. doi: 10.3390/ijms26178610.
Detailed analysis of gene expression by real time-quantitative polymerase chain reaction (RT-qPCR) has become a widespread method. To normalize the expression of target genes, this approach relies on constitutively expressed internal controls known as housekeeping genes (HKGs). Their proper selection is a critically important methodological step, since all the studied gene expression will be recalculated based on HKG expression. This concise review aims to discuss the selection of HKGs for endometrial cancer (EC) studies. We draw attention to the fact that the commonly used gene glyceraldehyde-3-phosphate dehydrogenase () is unsuitable as a HKG for research on the normal endometrium, EC, as well as many other tissues. In contrast, accumulating evidence suggests that GAPDH is a pan-cancer marker and an EC marker. Work on overexpression in EC in relation to overall and relapse-free survival is lacking. Both original research and overviews indicate that at least two HKGs should be used for target gene expression recalculations, a rarely applied technical aspect of final data processing. The insufficiently careful selection in many studies of only one HKG, e.g., , can be held responsible for broad discrepancies in published results obtained by this RT-qPCR technique. We provide an account of the discrepancies reported for sex hormone receptors expression in EC. Achieving consensus on the selection and validation of HKGs for research on this cancer is of crucial importance. Ideally, this trusted gene combination should be universal for any EC histotype and grade, irrespective of the final anatomopathological result.
通过实时定量聚合酶链反应（RT-qPCR）对基因表达进行详细分析已成为一种广泛应用的方法。为了使目标基因的表达标准化，这种方法依赖于组成性表达的内部对照，即管家基因（HKGs）。正确选择管家基因是一个至关重要的方法学步骤，因为所有研究的基因表达都将基于管家基因的表达进行重新计算。这篇简要综述旨在讨论子宫内膜癌（EC）研究中管家基因的选择。我们提请注意，常用的甘油醛-3-磷酸脱氢酶（）基因不适用于正常子宫内膜、子宫内膜癌以及许多其他组织的研究。相比之下，越来越多的证据表明GAPDH是一种泛癌标志物和子宫内膜癌标志物。关于GAPDH在子宫内膜癌中过表达与总生存期和无复发生存期关系的研究尚缺乏。原始研究和综述均表明，在重新计算目标基因表达时应至少使用两个管家基因，这是最终数据处理中一个很少应用的技术方面。在许多研究中，仅选择一个管家基因（例如）时不够谨慎，这可能是导致该RT-qPCR技术公布结果存在广泛差异的原因。我们阐述了在子宫内膜癌中报道的性激素受体表达差异。就子宫内膜癌研究中管家基因的选择和验证达成共识至关重要。理想情况下，这种可靠的基因组合对于任何子宫内膜癌组织学类型和分级都应是通用的，而不论最终的解剖病理学结果如何。
