title: Knockdown of ZNF403 inhibits cell proliferation and induces G2/M arrest by modulating cell-cycle mediators.
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.
authors.affiliation: 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@utoronto.ca
authors.forename: Rui
authors.initials: R
authors.lastname: Guan
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Wen
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Rongqi
authors.initials: R
authors.lastname: Duan
authors.forename: Huibi
authors.initials: H
authors.lastname: Cao
authors.forename: Simon
authors.initials: S
authors.lastname: Lam
authors.forename: Defu
authors.initials: D
authors.lastname: Hou
authors.forename: Youdong
authors.initials: Y
authors.lastname: Wang
authors.forename: Jim
authors.initials: J
authors.lastname: Hu
authors.forename: Zhuchu
authors.initials: Z
authors.lastname: Chen
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D002448:Cell Adhesion; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D059565:G2 Phase Cell Cycle Checkpoints; D015870:Gene Expression; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; C487856:GGNBP2 protein, human; D025521:Tumor Suppressor Proteins