Mechanism of prolactin inhibition of miR-135b via methylation in goat mammary epithelial cells.
Zhi ChenJun LuoChangHui ZhangYue MaShuang SunTianyin ZhangJuan J LoorPublished in: Journal of cellular physiology (2017)
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.
Keyphrases
- dna methylation
- dairy cows
- human milk
- poor prognosis
- growth hormone
- binding protein
- genome wide
- transcription factor
- low birth weight
- gene expression
- adipose tissue
- insulin resistance
- type diabetes
- long non coding rna
- single molecule
- cell proliferation
- skeletal muscle
- preterm infants
- atomic force microscopy
- preterm birth
- immune response
- cell free
- pi k akt
- data analysis
- toll like receptor
- circulating tumor cells
- light emitting