Effects of estradiol, progesterone, prolactin and oxytocin on miRNA levels in T47D human breast cells

The aim of the project was to analyse the levels of miRNA upon hormonal treatment in T47D breast cells T47D cells were obtained from ATCC and maintained in RPMI-1640 media without phenol red, supplemented with 10% fetal bovine serum (FBS). At 60 to 70% confluency, media was replaced with media containing hormone-stripped FBS for 16 hours. Then, cells were exposed to estradiol (10 nM, 24 hours), progesterone (10 nM, 6 hours), prolactin (8.7 nM (200 ng/mL), 8 hours) and oxytocin (100 nM, 24 hours for oxytocin) or the appropriate vehicle (0.1% ethanol for estrogen and progesterone, 4 mM hydrochloric acid for prolactin and nuclease-free water for oxytocin). miRNA were extracted and sequenced.