Long non-coding RNA FPFSC promotes immature porcine Sertoli cell growth through modulating the miR-326/EHMT2 axis

doi:10.1016/j.jia.2023.06.026

摘要/Abstract

摘要：

睾丸支持细胞是保证正常精子发生和雄性可育必不可少的细胞。尽管从不同发育时期的猪睾丸组织中鉴定出了大量的长链非编码RNA，并预测其在精子发生中具有关键的调控作用，但关于其功能和调控机制的研究仍处于初级阶段。因此，本研究旨在探究lncFPFSC在未成熟猪睾丸支持细胞增殖和凋亡中的功能及调控机理。结果表明，lncFPFSC主要表达于未成熟猪睾丸支持细胞的细胞质中，且过表达lncFPFSC促进细胞周期进程和细胞增殖并抑制细胞凋亡，而siRNA诱导的抑制表达lncFPFSC则具有相反的调控作用。机理研究表明，lncFPFSC作为“分子海绵”吸附抑制miR-326，过表达miR-326抑制细胞增殖并诱导细胞凋亡，且能够拮抗过表达lncFPFSC的作用。miR-326直接靶向常染色组蛋白赖氨酸N-甲基转移酶2基因，并抑制其mRNA和蛋白表达水平。此外，siRNA诱导的抑制表达EHMT2的作用与抑制表达miR-326的一致。综上，lncFPFSC通过调节miR-326/EHMT2信号轴促进未成熟猪睾丸支持细胞的增殖并抑制其凋亡。本研究拓展了我们对长链非编码RNA通过决定睾丸支持细胞的命运参与猪精子生成过程的理解，且该竞争性内源RNA网络为治疗支持细胞疾病诱导的雄性不育提供了新的分子标记。

Abstract:

Sertoli cells are indispensable for guaranteeing normal spermatogenesis and male fertility. Although a huge number of long non-coding RNAs (lncRNAs) are identified from developing porcine testicular tissues and have been predicted with crucial regulatory roles in spermatogenesis, their functions and regulatory mechanisms are still in infancy. Herein, we mainly explored the regulatory and functional roles of lncFPFSC in proliferation and apoptosis of immature porcine Sertoli cells. The results demonstrated that lncFPFSC was predominantly located in the cytoplasm of immature porcine Sertoli cells. lncFPFSC overexpression promoted cell cycle progression and cell proliferation, as well as inhibited cell apoptosis, whereas siRNA-induced lncFPFSC knockdown resulted in the opposite effects. Mechanistically, lncFPFSC acted as a sponge for miR-326. Overexpression of miR-326 inhibited cell proliferation and induced cell apoptosis, which further abolished the effects of lncFPFSC overexpression. The euchromatic histone-lysine N-methyltransferase 2 (EHMT2) gene was directly targeted by miR-326, and its mRNA and protein expressions were both negatively regulated by miR-326 in immature porcine Sertoli cells. Then, siRNA-induced EHMT2 knockdown resulted a similar effect of miR-326 inhibition. Collectively, lncFPFSC promoted proliferation and inhibited apoptosis in immature porcine Sertoli cells through modulating the miR-326/EHMT2 axis. This study expanded our understanding of non-coding RNAs in participating porcine spermatogenesis through deciding the fate of Sertoli cells, and the competing endogenous RNA (ceRNA) network, and provided new molecular markers to treat Sertoli cell disorder inducing male infertility.

Key words: porcine Sertoli cells , proliferation , apoptosis , lncRNA FPFSC , miR-326 , EHMT2 gene

Dan Chu, Bin Chen, Bo Weng, Saina Yan, Yanfei Yin, Xiangwei Tang, Maoliang Ran. [J]. Journal of Integrative Agriculture, 2024, 23(11): 3830-3842.

Dan Chu, Bin Chen, Bo Weng, Saina Yan, Yanfei Yin, Xiangwei Tang, Maoliang Ran. Long non-coding RNA FPFSC promotes immature porcine Sertoli cell growth through modulating the miR-326/EHMT2 axis[J]. Journal of Integrative Agriculture, 2024, 23(11): 3830-3842.

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