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鸡 GHR 反义转录本在肝细胞中调节其有义转录本。

Chicken GHR antisense transcript regulates its sense transcript in hepatocytes.

机构信息

Agricultural College, GuangDong Ocean University, ZhanJiang, China.

Guangdong Provincial Key Lab of Agro-animal Genomics and Molecular Breeding and Key Lab of Chicken Genetics, Breeding and Reproduction, Ministry of Agriculture, South China Agricultural University, Guangzhou, China.

出版信息

Gene. 2019 Jan 15;682:101-110. doi: 10.1016/j.gene.2018.10.001. Epub 2018 Oct 5.

Abstract

An increasing number of evidences indicated that long noncoding RNAs (LncRNAs) regulate a variety of biological progresses via different mechanisms. Our previous study had identified a chicken growth hormone receptor (GHR) antisense transcript (GHR-AS) which regulated GHR sense transcript (GHR-S) in LMH cells. In the present study, roles of GHR-AS and its regulatory mechanism were analyzed in chicken hepatocytes. The expression patterns of liver GHR-S, GHR-AS and Let-7b ascended with the development of chicken. The hepatocytes proliferation was promoted and more cells entered into DNA synthesis (S) phase when GHR-AS was overexpressed while the cell proliferation was slowed and fewer cells were in S phase when GHR-AS was interfered. Meanwhile, the GHR-S increased when we overexpressed GHR-AS while it reduced when GHR-AS was inhibited. The S1 Nuclease protection assay indicated that GHR-S and GHR-AS formed RNA duplex via GHR-S 3' untranslation regon (3'UTR). In hepatocytes or LMH cells, the half-time of GHR-S showed a delayed trend when GHR-AS or GHR-AS 5' untranslation regon (5'UTR) was overexpressed. Furthermore, the level of GHR-S can be decreased by Let-7b mimics whereas it was partially rescued when co-transfected pGHR-AS or pGHR-AS 5'UTR with Let-7b mimics. Based on our findings, GHR-AS affected hepatocytes proliferation and improved GHR-S stability possibly by forming RNA duplex between GHR-S and GHR-AS, competing with Let-7b.

摘要

越来越多的证据表明,长非编码 RNA(LncRNA)通过不同的机制调节多种生物学进程。我们之前的研究已经鉴定出一种鸡生长激素受体(GHR)反义转录本(GHR-AS),它在 LMH 细胞中调节 GHR 有义转录本(GHR-S)。在本研究中,分析了鸡肝细胞中 GHR-AS 的作用及其调控机制。鸡肝脏 GHR-S、GHR-AS 和 Let-7b 的表达模式随着鸡的发育而上升。当 GHR-AS 过表达时,肝细胞增殖加快,更多的细胞进入 DNA 合成(S)期,而当 GHR-AS 被干扰时,细胞增殖减慢,进入 S 期的细胞减少。同时,当 GHR-AS 过表达时,GHR-S 增加,而当 GHR-AS 被抑制时,GHR-S 减少。S1 核酸酶保护试验表明,GHR-S 和 GHR-AS 通过 GHR-S 3'非翻译区(3'UTR)形成 RNA 双链。在肝细胞或 LMH 细胞中,当 GHR-AS 或 GHR-AS 5'非翻译区(5'UTR)过表达时,GHR-S 的半衰期呈现出延迟的趋势。此外,Let-7b 模拟物可以降低 GHR-S 的水平,而当与 Let-7b 模拟物共转染 pGHR-AS 或 pGHR-AS 5'UTR 时,GHR-S 的水平部分得到挽救。基于我们的发现,GHR-AS 通过形成 GHR-S 和 GHR-AS 之间的 RNA 双链,与 Let-7b 竞争,影响肝细胞增殖并提高 GHR-S 的稳定性。

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