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Knockdown of ZNF403 inhibits cell proliferation and induces G2/M arrest by modulating cell-cycle mediators

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Abstract

ZNF403, also known as GGNBP2 (gametogenetin binding protein 2), is a highly conserved gene implicated in spermatogenesis. However, the exact biological function of ZNF403 is not clear. In this study, we identified the role of ZNF403 in cell proliferation and cell-cycle regulation by utilizing short hairpin RNA (shRNA)-mediated knockdown. ZNF403-specific shRNA expressing helper-dependent adenoviral vector (HD-Ad-ZNF403-shRNA) was constructed and transduced human cell lines. ZNF403 mRNA and protein expression levels were inhibited as evidenced by real-time PCR and western blot analyses. Noticeably, we found that knockdown of ZNF403 expression suppressed cell proliferation compared to the non-target shRNA and vector controls. Furthermore, cell-cycle analysis demonstrated that downregulation of ZNF403 promoted G2/M cell-cycle arrest in a dose-dependent manner. Moreover, human cell-cycle real-time PCR array revealed that ZNF403 knockdown influenced the expression profile of genes in cell-cycle regulation. Among these genes, western blot analysis confirmed the protein up-regulation of p21 and down-regulation of MCM2 in response to the ZNF403 knockdown. Additionally, knockdown of ZNF403 also showed an anti-carcinogenetic effect on anchorage-independent growth by colony formation assay and tumor cell migration by wound-healing assay with human laryngeal cancer cell line Hep-2 cells. Altogether, our findings suggest an essential role of ZNF403 in cell proliferation and provide a new insight into the function of ZNF403 in regulating the G2/M cell-cycle transition.

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Acknowledgments

This study was partially supported by grants from the Canadian Institutes of Health Research to JH and Heart and Stroke Foundation of Canada to XYW. RG is an awardee of the Chinese Scholarship Council (CSC).

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None of the authors has any type of conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital and Cancer Research Institute, Central South University, Changsha, 410008, Hunan Province, China

    Rui Guan, Defu Hou & Zhuchu Chen

  2. Physiology and Experimental Medicine Program, Hospital for Sick Children, 555 University Avenue, Toronto, ON, M5G 1X8, Canada

    Rui Guan, Jing Wu, Rongqi Duan, Huibi Cao, Simon Lam & Jim Hu

  3. Keenan Research Center, Li Ka Shing Knowledge Institute, St. Michael’s Hospital and Department of Medicine, University of Toronto, Toronto, ON, Canada

    Rui Guan, Xiao-Yan Wen & Youdong Wang

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  1. Rui Guan
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Correspondence to Jim Hu or Zhuchu Chen.

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Guan, R., Wen, XY., Wu, J. et al. Knockdown of ZNF403 inhibits cell proliferation and induces G2/M arrest by modulating cell-cycle mediators. Mol Cell Biochem 365, 211–222 (2012). https://doi.org/10.1007/s11010-012-1262-6

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  • DOI: https://doi.org/10.1007/s11010-012-1262-6

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