Vega-Sanchez R, Arenas-Hernandez M, Vazquez-Perez J A, Moreno-Valencia Y, Gomez-Lopez N
Department of Nutrition and Bioprogramming, Instituto Nacional de Perinatología, Montes Urales 800, México D.F. 11000, Mexico.
Department of Obstetrics & Gynecology, School of Medicine, Wayne State University, Detroit, MI, USA; National School of Biological Sciences, Instituto Politécnico Nacional, México D.F., Mexico.
Placenta. 2015 Feb;36(2):240-5. doi: 10.1016/j.placenta.2014.11.010. Epub 2014 Nov 26.
Human labor is considered an inflammatory process modulated by systemic and local leukocytes that infiltrate into the maternal-fetal interface. The putative roles of these leukocytes are currently being studied with gene expression assays. Such assays are normalized by the expression of housekeeping genes. However, expression of housekeeping genes may vary depending on the cell type and/or the experimental conditions. The aim of this study was to analyze the expression stability of several housekeeping genes in leukocytes from term human pregnancies, considering both anatomical origin and presence of labor.
We analyzed the gene expression of ACTB, B2M, GAPDH, GUSB, PGK1, RN18S1, TBP and UBC in leukocytes from maternal peripheral blood, placental blood and choriodecidua in women delivering at term with or without the presence of labor through real-time qPCR. Then we used geNorm to evaluate expression stability and pairwise variation.
The expression of all tested genes showed to be stable independent of the anatomical compartment and the absence or presence of labor. However, PGK1, GUSB and TBP showed to be the most stable and RN18S1 the least stable. Pairwise variation analyses showed that only two genes are needed for normalization yet the inclusion of a third improves its accuracy.
PGK1, GUSB and TBP are the most adequate reference genes for gene expression normalization in leukocytes from term pregnancies, regardless of their anatomical origin (maternal peripheral blood, placental blood or choriodecidua) or the presence or absence of labor. Our study is the first report on housekeeping gene stability in leukocytes from healthy pregnant women.
人类分娩被认为是一个由浸润至母胎界面的全身和局部白细胞调节的炎症过程。目前正在通过基因表达分析研究这些白细胞的假定作用。此类分析通过管家基因的表达进行标准化。然而，管家基因的表达可能因细胞类型和/或实验条件而异。本研究的目的是分析足月妊娠白细胞中几个管家基因的表达稳定性，同时考虑解剖学来源和分娩情况。
我们通过实时定量PCR分析了足月分娩或未分娩女性的外周血、胎盘血和绒毛膜蜕膜白细胞中ACTB、B2M、GAPDH、GUSB、PGK1、RN18S1、TBP和UBC的基因表达。然后我们使用geNorm评估表达稳定性和成对变异。
所有测试基因的表达均显示稳定，与解剖部位以及是否存在分娩无关。然而，PGK1、GUSB和TBP显示最稳定，而RN18S1最不稳定。成对变异分析表明，标准化仅需要两个基因，但加入第三个基因可提高其准确性。
PGK1、GUSB和TBP是足月妊娠白细胞基因表达标准化的最合适参考基因，无论其解剖学来源（母体外周血、胎盘血或绒毛膜蜕膜）以及是否存在分娩。我们的研究是关于健康孕妇白细胞中管家基因稳定性的首次报告。
