Ovarian IL-16 and miR-125a-5p Expression

Expression of IL-16 gene and its regulator microRNA miR-125a-5p was examined in representative specimens (8 premenopausal ovaries, 6 early menopausal ovaries, and 4 late menopausal ovaries) by quantitative real-time polymerase chain reaction (qRT-PCR). Total RNA was extracted from all specimens using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's recommendation. RNA was then measured at an optical density (OD) of 260 nm and an OD of 260/280 nm absorbance ratio ≥1.7 was used to evaluate the purity, as previously reported [30 (link)].
The expression of IL-16 messenger RNA (mRNA) and miR-125a-5p in normal ovaries and fimbriae was measured by quantitative real-time polymerase chain reaction (qRT-PCR). The human specific IL-16 primer (QT00075138) designed by QuantiTech and miR-125a-5p designed by Applied Biosystems (Foster City, CA) were used for qRT-PCR analyses. β-Actin was used as housekeeping gene in qRT-PCR experiments. Gene expression amplification was determined using the method of the differences (δ) in cycle threshold (ΔCt) for IL-16 mRNA and miR-125a-5p according to the manufacturer's recommendation. Subtracting ΔCt from each group from the average ΔCt determined the ΔΔCt. 2^–ΔΔCt was used to calculate the fold change in the differences in IL-16 mRNA and miR-125a-5p expression levels.