Mechanism of prolactin inhibition of miR-135b via methylation in goat mammary epithelial cells

Zhi Chen, Jun Luo, Chang Hui Zhang, Yue Ma, Shuang Sun, Tianyin Zhang, Juan J. Loor

Research output: Contribution to journalArticle

Abstract

Prolactin is an important endocrine activator of lactogenesis. This study investigated the function and mechanism of miR-135b in the enhancement of lactation by prolactin in goat mammary epithelial tissue. We utilized S-Poly (T) sequencing to evaluate changes in gene regulation in the goat mammary gland after incubation with 2.5 μg/ml prolactin and 2.5 μg/ml IGF-1 by examining highly expressed miRNAs during early lactation and late-lactation. The results illustrated that miR-135b is highly expressed in the goat mammary gland during early lactation and late-lactation, and also after treatment with 2.5 μg/ml prolactin and 2.5 μg/ml IGF-1. We used Q-RT PCR, Western Blot, immunofluorescence, and luciferase reporter assay analysis, and found that PRL was significantly down-regulated in response to the expression of miR-135b in a manner that was functionally related to TAG synthesis via the large tumor suppressor 2 gene (Lats2), an important regulator of adipocyte proliferation via Hippo Signaling. Furthermore, using bisulfite-sequencing PCR (BSP), Q-PCR, and Western Blot we discovered an increase in expression of DNMT I (DNA methyl transferase I) in goat mammary epithelial cells with the 2.5 μg/ml PRL incubation, which led to DNA methylation of the CpG island upstream of miR-135b and inhibited the transcription and expression of miR-135b.

Original languageEnglish (US)
Pages (from-to)651-662
Number of pages12
JournalJournal of Cellular Physiology
Volume233
Issue number1
DOIs
StatePublished - Jan 2018

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Keywords

  • LATS2
  • fat metabolism
  • methylation
  • miR-135b

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

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