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LncRNA HOTAIR promotes proliferation and suppresses apoptosis of mouse spermatogonium GC-1 cells by sponging miR-761 to modulate NANOS2 expression

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Abstract

LncRNA HOX antisense intergenic RNA (HOTAIR) can regulate cancer-related gene expression and promote stem cell and tumor cell proliferation via mechanisms including the competing endogenous RNA (ceRNA) mechanism. HOTAIR is abundantly expressed in the genital tubercle of E11.5, E12.5, and E13.5 embryos, whereas it became barely detectable at E13.5 and expressed again in adult mouse testis. However, the underlying function and mechanism of HOTAIR in spermatogenesis have not been elucidated. Interestingly, other researchers reported that the function of gene Nanos C2HC-Type Zinc Finger 2 (nanos2) includes the maintenance of both the primordial germ cells (PGCs) and germline stem cells, and Nanos2 protein and transcripts (NANOS2) were detected only in PGCs from day E11.5 and undifferentiated spermatogonia in spermatogenesis. We therefore investigated the relationship between HOTAIR and NANOS2 in maintaining spermatogonial stem cell population. We found that, compared to the adult mouse, the expression levels of HOTAIR and NANOS2 in embryo mouse were significantly higher and miR-761expression level was lower. In mouse GC-1 spermatogonia cells, overexpression of miRNA-761 significantly inhibited the expression of NANOS2 and HOTAIR, suppressed the proliferation, and promotes apoptosis of cells. Knock down and overexpression of HOTAIR indicated that HOTAIR expression was positively correlated with NANOS2 expression; overexpressed HOTAIR could promote proliferation and suppresses apoptosis of GC-1 cells. By a rescue experiment and dual luciferase reporter assay, miR-761 was identified as a direct target of HOTAIR, and NANOS2 was identified as the direct target of miR-761. The above results indicate that HOTAIR promotes proliferation and suppresses apoptosis of mouse spermatogonium GC-1 cells by sponging miR-761 to modulate NANOS2 expression. Our findings elucidate one of possible mechanisms and importance of HOTAIR in maintaining spermatogonial stem cell population, and provide new candidate genes and possible pathogenesis for male infertility.

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Data availability

The data that support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

We sincerely thank all members of the Department of Reproductive Medical Center at the First Affiliated Hospital of Wenzhou Medical University.

Funding

This work was supported by the Science and Technology Bureau of Wenzhou, China (Grant Y20180065) and the National Key Research and Development Program of China (2017YFC1002001).

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Authors and Affiliations

  1. Department of Reproductive Medical Center, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, China

    Xiangbin Kong, Qianjin Fei, Chengshuang Pan, Jianyuan Jin, Jiujia Zheng & Xuefeng Huang

  2. Wuhan GeneCreate Biological Engineering Co., Ltd., Wuhan, 430000, China

    Didi Wu

  3. Institute of Reproductive Health/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China

    Honggang Li

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  1. Xiangbin Kong
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Contributions

Xiangbin Kong designed the study and its analytic strategy and wrote the original draft. Didi Wu assisted in bioinformatic analysis and technical guidance. Qianjin Fei, Chengshuang Pan, Jianyuan Jin, Jiujia Zheng, and Didi Wu performed the research. Honggang Li and Xuefeng Huang designed the research study and edited the original draft. All authors read and approved the final manuscript.

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Correspondence to Honggang Li or Xuefeng Huang.

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Kong, X., Fei, Q., Pan, C. et al. LncRNA HOTAIR promotes proliferation and suppresses apoptosis of mouse spermatogonium GC-1 cells by sponging miR-761 to modulate NANOS2 expression. In Vitro Cell.Dev.Biol.-Animal 58, 295–306 (2022). https://doi.org/10.1007/s11626-022-00657-y

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