Testis-enriched circular RNA circ-Bbs9 plays an important role in Leydig cell proliferation by regulating a CyclinD2-dependent pathway
Testis-enriched circular RNA circ-Bbs9 plays an important role in Leydig cell proliferation by regulating a CyclinD2-dependent pathway
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富含睾丸的环状 RNA circ-Bbs9 通过调节 CyclinD2 依赖性途径在 Leydig 细胞增殖中发挥重要作用
DOI：
10.1071/rd18474
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Xu Wenming
中科院分区：
文献类型：
--
作者：
Jia Minzhi;Li Xiaoliang;Jiang Chuan;Wang Ke;Zuo Tao;He Guolin;Qin Lang;Xu Wenming
环状RNA是一类新的非编码RNA，具有环状结构、保守性、稳定性和在真核生物中的高表达等特点。它们通常显示组织或细胞特异性表达。在这里，我们确定了一个睾丸富集的环状RNA（circRNA），环状Bbs 9（circ-Bbs 9），它在小鼠睾丸中高度表达。RNase R处理实验证实了circ-Bbs 9确实是一种circRNA。原位杂交实验表明，circ-Bbs 9表达在睾丸间质细胞沿着曲细精管和在细胞质中的TM 3睾丸间质细胞系。慢病毒载体敲低circ-Bbs 9基因可抑制TM 3细胞增殖，而过表达circ-Bbs 9则可显著诱导TM 3细胞增殖。敲低circ-Bbs 9可抑制cyclin D2（Ccnd 2）的蛋白水平，RNA免疫沉淀结果显示circ-Bbs 9与Ccnd 2相互作用。我们的研究结果表明，使用刺猬途径Smoothened激动剂（SAG）HCl和拮抗剂cyclopamine和gant 6影响神经胶质瘤相关癌基因同源物1（Gli 1），Ccnd 2和其他基因在这条途径中的表达水平。我们的研究表明，间质细胞特异性circRNA，circ-Bbs 9，通过调节细胞周期相关的Ccnd 2的水平在间质细胞增殖中起着关键作用。因此，我们的研究结果强调了circRNA在男性生殖系统中的重要作用。
Circular RNAs belong to a new category of non-coding RNAs, characterised by a circular structure, conservation, stability and high expression in eukaryotes. They often show tissue- or cell-specific expression. Here, we identified a testis-enriched circular RNA (circRNA), circular Bbs9 (circ-Bbs9) that is highly expressed in mouse testis. An RNase R treatment experiment confirmed that circ-Bbs9 is indeed a circRNA. In situ hybridisation experiments showed that circ-Bbs9 is expressed in Leydig cells along seminiferous tubules and in the cytoplasm of the TM3 Leydig cell line. Knocking down the circ-Bbs9 in TM3 cells by lentivirus vectors arrested cell proliferation, whereas overexpression of circ-Bbs9 induced cell proliferation significantly. Knocking down circ-Bbs9 inhibited the protein level of cyclin D2 (Ccnd2) and RNA immunoprecipitation results showed that circ-Bbs9 interacts with Ccnd2. Our results show that use of the Hedgehog pathway Smoothened Agonist (SAG) HCl and antagonists cyclopamine and gant6 affects the expression levels of Glioma-Associated Oncogene Homolog 1 (Gli1), Ccnd2 and other genes in this pathway. Our research reveals that a Leydig cell-specific circRNA, circ-Bbs9, plays a critical role in Leydig cell proliferation through regulating the levels of cell cycle-related Ccnd2. Thus, our results emphasise the important role of circRNA in the male reproductive system.