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MIR-182 AIDS in receptive endometrium development in dairy goats by downregulating PTN expression

PLoS ONE (2017) 12(7)
DOI: 10.1371/journal.pone.0179783
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Abstract

Increasing evidence has shown that miRNAs play important roles in endometrium development during the menstrual cycle in humans and many other animals. Our previous data indicated that miR-182 levels increase 15.55-fold and pleiotrophin (PTN) levels decrease 20.97-fold in the receptive endometrium (RE, D15) compared with the pre-receptive endometrium (PE, D5) in dairy goats. The present study shows that miR-182 is widely expressed in different tissues of dairy goats and that its expression levels are regulated by E2 and P4 in endometrial epithelium cells (EECs). We confirmed that PTN is a target of miR-182 and that miR-182 regulates the protein levels of AKT, Bcl-2, FAS, MAPK, Caspase-3 and SP1 in EECs. Furthermore, miR-182 up-regulates or maintains the expression levels of osteopontin (OPN), cyclooxygenase-2 (COX-2) and prolactin receptor (PRLR) in EECs, suggesting that miR-182 is an important regulatory factor in the construction of endometrial receptivity in dairy goats. In conclusion, miR-182 participates in the development of endometrial receptivity by down-regulating PTN and affecting the expression of select apoptosis-related genes and increasing or maintaining the expression levels of OPN, COX-2 and PRLR in the EECs of dairy goats.

Figures

  • Table 1. The RT-qPCR Primers used in the present study.
  • Table 2. The antibody used in the present study.
  • Fig 1. Expression levels of miR-182 in endometrium at D5 and D15. The miR-182 levels in endometrium at D5 and D15 were detected by stem-loop qRT-PCR, 18S rRNA (A) and U6 (B) were used as the references. The values were showed as “means ± SD” (n = 5×3), ** indicates that P < 0.01.
  • Fig 2. miR-182 was expressed in various tissues of dairy goats. The miR-182 levels in different tissues were measured by Stem-loop RT-qPCR and normalized to 18S. Values were showed as “means ± SD” in receptive endometrium (RE, D15) that were relative to pre-receptive endometrium (PE, D5); n = 5×3. ** indicates that P < 0.01 and * indicates that P < 0.05.
  • Fig 3. The effect of E2 and P4 on the expression levels of miR-182 in endometrial epithelium cells (EECs). The effect of E2 and P4 on the expression levels of miR-182 in EEC were measured by Stem-loop RT-qPCR and normalized to 18S. Values were showed as “means ± SD” (n = 3×3). * mean that the difference was significant compared to the control cells (did not added E2 and P4 in cell medium).
  • Fig 4. The effect of miR-182 on the surface microtopography of endometrial epithelium cells (EECs).
  • Fig 5. The effect of miR-182 on the proliferation of endometrial epithelium cells (EECs). The effect of miR-182 on the proliferation of EEC was measured by MTT. Values were showed as “means ± SD” (n = 3×7). ** indicates that P < 0.01 and * indicates that P < 0.05.
  • Fig 6. The effect of miR-182 on the cell cycle of endometrial epithelium cells (EECs). The cell cycle analysis of EEC were made after the cells were transfected with miR-182 mimic (A); NC (B); miR-182 inhibitor (C); NC inhibitor (D).

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Zhang, L., Liu, X., Liu, J., Zhou, Z., Song, Y., Cao, B., & An, X. (2017). MIR-182 AIDS in receptive endometrium development in dairy goats by downregulating PTN expression. PLoS ONE, 12(7). https://doi.org/10.1371/journal.pone.0179783

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