期刊
JOURNAL OF CELLULAR PHYSIOLOGY
卷 233, 期 1, 页码 651-662出版社
WILEY
DOI: 10.1002/jcp.25925
关键词
fat metabolism; LATS2; methylation; miR-135b
资金
- Special Fund for Agro-scientific Research in the Public Interest [201103038]
- Transgenic New Species Breeding Program of China [2014ZX08009-051B]
- National Natural Science Foundation of China [31372281]
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 mu g/ml prolactin and 2.5 mu g/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 mu g/ml prolactin and 2.5 mu g/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 mu g/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.
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