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首页> 外文期刊>Journal of Cellular Physiology >Mechanism of prolactin inhibition of miR‐135b via methylation in goat mammary epithelial cells
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Mechanism of prolactin inhibition of miR‐135b via methylation in goat mammary epithelial cells

机译:催乳素抑制机制米尔135 b通过甲基化在山羊乳腺上皮细胞

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摘要

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.
机译:催乳素是一种重要的内分泌激活生乳。米尔135 b应承担的功能和机制催乳激素增强哺乳的山羊乳腺上皮组织。测序来评估基因调控的变化在孵化后的山羊乳腺2.5 ?检查高度表示microrna在早期哺乳期和后期的哺乳。说明米尔135 b是高度表达在早期的哺乳和山羊乳腺治疗后晚还是哺乳,也2.5 ?问量RT PCR,免疫印迹、免疫荧光和荧光素酶报告实验分析,发现, PRL必经的监管应对米尔量135 b的表达功能相关的标记方式通过2大肿瘤抑制基因合成(Lats2),脂肪细胞的重要调节器通过河马信号扩散。用亚硫酸氢量测序PCR (BSP),问高PCR和免疫印迹我们发现增加DNA甲基转移酶(DNMT表达我)在山羊乳腺上皮细胞2.5 ?甲基化的CpG岛上游米尔135 b和抑制转录表达米尔135 b。

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