mRNA-Expression of Often Used House-Keeping Genes and the Relation between RNA and DNA Are Sex Steroid-Dependent Parameters in Human Myometrium and Fibroids

Abstract

The content of RNA and DNA in human myometrium and fibroids obtained at different endocrine conditions varied, with the highest RNA/DNA ratio in tissues from pregnant patients, intermediate ratios in women during the menstrual cycle and the lowest in tissues from postmenopausal and GnRHa-treated patients. mRNA expression of two house-keeping genes, γ-actin and GAPDH, was highest in myometrium from pregnant women, intermediate in untreated women of fertile age and lowest in tissues from GnRHa-treated and postmenopausal women. To control for degradation of nucleic acids when measuring mRNA expression, we suggest additional analysis of gene(s), where the expression pattern is known, and that expression, whenever possible, is related to DNA, which is a more stable parameter than RNA and total nucleic acids, when there are differences in proliferation between tissues and/or groups of patients.

Development of a Macroarray To Specifically Analyze Immunological Gene Expression in Swine

“…In contrast, housekeeping genes are considered to be expressed constantly. However, several reports have indicated that the expression levels of these genes may change in different situations and tissues (11,14,15,22,32,35). In this paper, we demonstrated that the gene coding for HPRT was expressed uniformly among the tested samples and at the same level as the other immunological genes.…”

“…As a differential expression of housekeeping genes has been described recently (14,22,35), the expression levels of four putative housekeeping genes included in the array were determined. Compared to those of immunological genes, the levels of expression of three out of these four housekeeping genes were high: median arbitrary units were 58 ϫ 10 4 , 23 ϫ 10 4 , and 13 ϫ 10 4 for the ␤-actin, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and cyclophilin genes, respectively, and 2.1 ϫ 10 4 for the immunological genes.…”

Increased Expression of the Relaxin Receptor (LGR7) in Human Endometrium during the Secretory Phase of the Menstrual Cycle

“…A previous immunohistochemical study of endometrial sections from three patients showed little, if any, staining in sections from patients in the proliferative or early secretory phase, but intense staining in one patient in the mid-secretory phase. 12 Another study reported no consistent regulation of LGR7 mRNA during the phases of the menstrual cycle, 13 but the expression levels are expressed as a ratio with GAPDH, which is known to be sex-steroid dependent, 14 whereas in the present study quantitative PCR was carried out using 18S RNA as the standard. The previous study was also carried out with biopsy samples and not the full-thickness endometrial samples used here.…”

Detection of drug transporter expression using a 25-multiplex RT-PCR assay

“…As summarized in Table 2, the 25 genes for the multiplex RT-PCR assay included 17 ABC transporters (ABCA2, ABCA3, ABCB1, ABCB4, ABCB5, ABCC1, ABCC2, ABCC3, ABCC4, ABCC5, ABCC6, ABCC9, ABCC10, ABCC11, ABCG1, House keeping GAPDH GAPDHGlycolysis Wu et al (2003) ABCG2 and CFTR), one non-ABC transporter (MVP), 4 SLC transporters (SLC28A1, SLC28A2, SLC29A1, SLC29A2), 2 copper transporters (SLC31A1 and ATP7B) and one housekeeping gene (GAPDH). We designed the primers using the Primer3 program and set up combinations of 5 genes in each set.…”