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Journal of Integrative Agriculture  2024, Vol. 23 Issue (11): 3830-3842    DOI: 10.1016/j.jia.2023.06.026
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Long non-coding RNA FPFSC promotes immature porcine Sertoli cell growth through modulating the miR-326/EHMT2 axis
Dan Chu1, Bin Chen1#, Bo Weng1, 2, Saina Yan1, Yanfei Yin1, Xiangwei Tang1, Maoliang Ran1#

1 Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China

2  Xiangxi Vocational and Technical College for Nationalities, Xiangxi 416007, China

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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作用,但关于其功能和调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.


Keywords:  porcine Sertoli cells        proliferation        apoptosis        lncRNA FPFSC        miR-326        EHMT2 gene  
Received: 14 February 2023   Accepted: 16 May 2023
Fund: 

This research was supported by the special funds for Changsha Municipal Natural Science Foundation, China (kq2202229), the Hunan Provincial Natural Science Foundation of China (2023JJ30296 and 2023JJ60247), and a Key R&D Projects (2020NK2024) in Hunan Province, China (2020NK2024).

About author:  #Correspondence Maoliang Ran, E-mail: ranmaoliang0903@126.com; Bin Chen, E-mail: chenbin7586@126.com

Cite this article: 

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

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