Effect of inhibiting miR-29a expression on prolactin secretion of human endometrial stromal cells during in vitro decidualization

ZHANG Peng; JIANG Wanjun

doi:10.3969/j.issn.1002-266X.2016.07.001

Abstract：Objective To observe the effect of inhibiting miR-29a expression on prolactin secretion of human endom-etrial stromal cells ( HESCs) during in vitro decidualization.Methods We collected the endometrial tissues during pan-hysterectomy from patients with uterine benign lesions, and used trypsin method to isolate HESCs.The cells were then di-vided into the experimental group and control group, the experimental group was treated with estradiol ( E2 ) and progester-one (P4), and the control group was not treated.We took part of the cells in 2 ×105/mL to inoculate into the 6-well plates and and then divided them into groups1, 2 and 3.The group 1 was transfected by anti-miR-29a.Group 2 was transfected by Control-miRNA and group 3 as taken as the blank control.After being cultured for 0 h, 24 h, 48 h, 72 h we collected cells in each group.real-time PCR was used to detect miR-29a in the cells of each group.dPRL in the supernatant of cell culture medium was detected by chemiluminescence method.Results The relative expression of miR-29a in the cells after being cultured for 0 h, 24 h, 48 h, 72 h, and the dPRL level in the cell culture supernatantthe of the experimental group were higher than those of the control group (all P<0.01).In the experimental group, with the prolonged culture time, the miR-29a expression was gradually increased, and the level of dPRL in the culture supernatant was increased.Significant difference was found between every two time points (all P<0.05).The relative expression of miR-29a in the group 1 at 24 h, 48 h and 72 h was lower than that in the groups 2 and 3, and the level of dPRL in the supernatant of group 1 was lower than that in the groups 2 and 3 (all P<0.01).Conclusions After the inhibition of miR-29a expression, human decidual endometrial stromal cells reduce the secretion of dPRL.miR-29a may be involved in the decidual process of HESCs, and regulates the endometrial stromal cells secreting dPRL.