Let-7g-5p Regulates Mouse Mammary Cells Differentiation and Function by Targeting PRKCA
Let-7g-5p Regulates Mouse Mammary Cells Differentiation and Function by Targeting PRKCA
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Let-7g-5p 通过靶向 PRKCA 调节小鼠乳腺细胞的分化和功能
DOI：
10.1002/jcp.27676
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Li Q
中科院分区：
文献类型：
--
作者：
Tian L;Li Y;Wang C;Li Q
MicroRNAs(MiRNAs)与基因表达的转录后调控密切相关。我们既往的研究表明，let-7g-5p在小鼠乳腺中的表达在妊娠和哺乳期均呈下降趋势，提示let-7g-5p对乳腺上皮细胞有潜在的调控作用。在本研究中，为了揭示let-7g-5p的功能，我们通过生物信息学的研究发现，PrKca(PKC-α，PKC-α)可能是let-7g-5p的潜在靶标。然后用反义寡核苷酸将let-7G-5p在小鼠原代乳腺上皮细胞中击倒。不出所料，在体外，抗let-7G-5p调控可促进PRKCA基因的表达和乳腺上皮细胞的生长。利用报告构建的方法表明，let-7G-5p可以直接与PRKCA基因的3‘-非翻译区(3’-UTR)结合。此外，体内异位过表达let-7g-5p降低了PRKCA和β-酪蛋白的表达，并抑制了乳腺生长。这些结果提示，let-7g-5p可能通过抑制乳腺细胞分化发育周期而抑制乳腺上皮细胞分化和β-酪蛋白蛋白的合成和表达，let-7g-5p可能是乳腺细胞功能的重要调控靶点。
MicroRNAs (miRNAs) are closely related with the posttranscriptional regulation of gene expression. Our past study showed that let‐7g‐5p decreased during pregnancy and lactation in mouse mammary gland, what suggested the prospective regulating role of let‐7g‐5p in mammary epithelial cells. In this study, to unravel the role of let‐7g‐5p, we found PRKCA (PKC‐alpha, PKC‐α) might serve as potential targets of let‐7g‐5p by bioinformatics. Then let‐7g‐5p was knocked down by an antisense oligonucleotide in mouse primary mammary epithelial cells. As predicted,PRKCAgene expression and mammary epithelial cells growth were increased by anti‐let‐7g‐5p regulation in vitro. By using reporter constructs, it showed that the let‐7g‐5p could direct binding with 3′‐the untranslated regions (3′‐UTR) of PRKCA mRNA. Furthermore, ectopic overexpression of let‐7g‐5p in vivo reduced PRKCA and β‐casein protein expression as well as inhibited mammary gland growth. These results suggested that let‐7g‐5p could inhibit the mammary epithelial cells differentiation and β‐casein protein synthesis and expression through suppression of PRKCA on the cycle of mammary cell differentiation and development and that let‐7g‐5p may be a novel important regulated target in mammary cells function.