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Molecular Biology Reports

, Volume 46, Issue 2, pp 2029–2038 | Cite as

Cloning of a new testis-enriched gene C4orf22 and its role in cell cycle and apoptosis in mouse spermatogenic cells

  • Hui Li
  • Yanfa Dai
  • Zhongqin LuoEmail author
  • Dongsong NieEmail author
Original Article
  • 122 Downloads

Abstract

Spermatogenesis is a complicated and dynamic cellular differentiation process mainly regulated by genes, steroid hormones and environmental factors. Although a number of genes involved in spermatogenesis have been identified, there are still a lot of genes underlying spermatogenesis remained unexplained. Here, a novel gene C4orf22, also known as 1700007G11Rik or Cfap299 was identified from mouse testis. C4orf22 protein contains 233 amino acid residues and is highly conserved in metazoan species. C4orf22 mRNA was predominantly expressed in mouse testis and increased from 2-week-old testes to 8-week-old testes during the developing testes by RT-PCR and qRT-PCR. Immunohistochemical analysis indicated that C4orf22 protein was mainly distributed in the cytoplasm of spermatogonia and primary spermatocytes, which was further confirmed by C4orf22-tagged with GFP in the GC-1 and GC-2 cells. Over-expression of pEGFP-C3-C4orf22 significantly inhibited GC-1 cells apoptosis and promoted cell cycle progression with an increase in the cell number of S and G2 phase. Conversely, small interfering RNA (siRNA) silencing C4orf22 in GC-1 cells could cause an increase in the number of apoptosis cells and the cell cycle was arrested at G2/M phase. Western blot analysis and qRT-PCR results showed that C4orf22 over-expression significantly increased the expressions of anti-apoptotic bcl-2 and decreased the expression of caspase-3, caspase-8 and Bax. Our results suggest that C4orf22 may be involved in spermatogenesis, and for the first time, unravels its potential role in regulating cell apoptosis through bcl-2 regulatory pathway in GC-1 cells.

Keywords

C4orf22 gene Gene expression Cell cycle Apoptosis 

Notes

Acknowledgements

This study was supported by the National Natural Science Foundation of China (Nos. 31571277, 30971570) and Hunan Provincial Natural Science foundation of China (2017JJ2104 and 2017JJ2109).

Author contributions

Conceived and designed the experiments: DN; Performed the experiments: HL and DY; Contributed reagents/materials/analysis tools: DN, HL and LZ; Wrote the paper: HL and DN; Completed calculations and participated in the interpretation of the results: HL.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All animal handling complied with the requirements of the directory of the Ethical Treatment of Experimental Animals of China.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Chemistry and Chemical EngineeringHunan Institute of Science and TechnologyYueyangPeople’s Republic of China

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