Role of micro RNA on the Expression of Luteinizing Hormone-Human Chorionic Gonadotropin Receptor Messenger Ribonucleic Acid in rat ovary

微小RNA对大鼠卵巢黄体生成素-人绒毛膜促性腺激素受体信使核糖核酸表达的影响

基本信息

批准号：
21390448
负责人：
MINEGISHI Takashi
金额：
$ 11.32万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2011
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21390448/
关键词：
miRNA LH receptor down regulation 卵巣 microRNA micro RNA シグナル伝達

项目摘要

MicroRNAs (miRNAs) are small non-coding RNAs that interact with mRNAs and trigger either translation repression or RNA cleavage of target genes. In this study, we investigated whether miRNA is involved in down-regulation of the LH receptor (LHR) in the ovary. A miRNA microarray was carried out to analyze the overall miRNA expression profile while LHR mRNA was down-regulated, and found that 23 miRNAs were highly expressed during this period. Combining these results with data from the bioinformative database, the clustering analysis led us to focus on miR-136* for further analysis. In both in vivo and in vitro studies, miR-136* levels were found to increase 6 hr after hCG administration while LHR mRNA levels were down-regulated. To confirm that miR-136* binds to LHR mRNA, we cloned the 3'-end of LHR mRNA from 2570 to 2895 into reporter vector which contained a Renilla luciferase coding region upstream of the cloning site. When the miR-136* inhibitor was co-transfected with the reporter vector in granulosa cells, the luciferase activity was de-repressed, revealing that miR-136* definitely bound to the 3'-end of LHR mRNA. From these data, we conclude that miR-136* participates in the mechanism of down-regulation of LHR mRNA, whereby miR-136* forms base pairs with LHR mRNA.

MicroRNA (miRNA) 是一种小型非编码 RNA，可与 mRNA 相互作用并触发靶基因的翻译抑制或 RNA 裂解。在这项研究中，我们研究了 miRNA 是否参与卵巢 LH 受体 (LHR) 的下调。通过miRNA微阵列分析LHR mRNA下调期间的整体miRNA表达谱，发现23个miRNA在此期间高表达。将这些结果与生物信息数据库中的数据相结合，聚类分析使我们重点关注 miR-136* 进行进一步分析。在体内和体外研究中，发现 hCG 给药后 6 小时，miR-136* 水平增加，而 LHR mRNA 水平下调。为了确认 miR-136* 与 LHR mRNA 结合，我们将 LHR mRNA 的 3' 端从 2570 克隆到 2895 到报告载体中，该报告载体在克隆位点上游含有海肾荧光素酶编码区。当 miR-136* 抑制剂与报告载体共转染颗粒细胞时，荧光素酶活性被解除抑制，表明 miR-136* 确实与 LHR mRNA 的 3' 端结合。从这些数据中，我们得出结论，miR-136* 参与 LHR mRNA 的下调机制，从而 miR-136* 与 LHR mRNA 形成碱基对。