Endometriosis and stem cells: identification of reference genes
Posted July 22, 2021
A pioneering study, developed by the INCT Hormona team at the Ribeirão Preto School of Medicine of Universidade de São Paulo (USP), identified the most suitable reference genes to be used in the quantification of the expression of messenger molecules in menstrual blood-derived stem cells from women with and without endometriosis.
According to professor and researcher Dr. Juliana Meola, the finding is of great importance in directing future studies. ‘The origin of endometriosis is enigmatic and has therefore been the subject of much investigation,’ she says. ‘Some ideas include that the endometrial tissue and, consequently, the cellular components present in menstrual blood are a first step toward the onset of the disease,’ she explains.
The expert states that, in recent decades, it has been suggested that stem cells present in the menstrual blood of women affected by the disease carry alterations at the level of messenger molecules (gene expression), thus contributing to the onset and development of endometriosis. ‘The gold standard technique for quantifying these molecules is reverse transcription-quantitative polymerase chain reaction (RT-qPCR), but the reliability of gene expression measurements using RT-qPCR is strongly affected by the use of housekeeping or unstable reference genes in data normalization calculations,’ says the expert.
In the study, the research group used stringent case and control selection criteria and collected menstrual blood from women with a laparoscopic diagnosis of advanced endometriosis and from fertile women without endometriosis. ‘We tested for the first time the stability of 32 candidate reference genes to achieve increased accuracy and reliable results in the quantification of gene expression and direct future experiments using RT-qPCR in menstrual blood-derived mesenchymal stem/stromal cells for endometriosis studies,’ she says. ‘Using the RefFinder web tool, we recommend the EIF2B1 and POP4 reference genes for the normalization of RT-qPCR data in study designs similar to ours and we suggest avoiding the commonly used GAPDH and ACTB reference genes as they are unstable,’ she warns.
Dr. Juliana Meola believes that the finding will contribute to future studies. ‘This study is capable of directing different experimental designs, in addition to endometriosis, as these stem cells are derived from a minimally invasive tissue source with multifunctional roles in regenerative medicine,’ she concludes.
The article “Identification of suitable reference genes for mesenchymal stem cells from menstrual blood of women with endometriosis” can be read in full at https://pubmed.ncbi.nlm.nih.gov/33686153/.