title: Transcriptional activation of human CDCA8 gene regulated by transcription factor NF-Y in embryonic stem cells and cancer cells.
abstract: The cell division cycle associated 8 (CDCA8) gene plays an important role in mitosis. Overexpression of CDCA8 was reported in some human cancers and is required for cancer growth and progression. We found CDCA8 expression was also high in human ES cells (hESCs) but dropped significantly upon hESC differentiation. However, the regulation of CDCA8 expression has not yet been studied. Here, we characterized the CDCA8 promoter and identified its cis-elements and transcription factors. Three transcription start sites were identified. Reporter gene assays revealed that the CDCA8 promoter was activated in hESCs and cancer cell lines. The promoter drove the reporter expression specifically to pluripotent cells during early mouse embryo development and to tumor tissues in tumor-bearing mice. These results indicate that CDCA8 is transcriptionally activated in hESCs and cancer cells. Mechanistically, two key activation elements, bound by transcription factor NF-Y and CREB1, respectively, were identified in the CDCA8 basic promoter by mutation analyses and electrophoretic motility shift assays. NF-Y binding is positively correlated with promoter activities in different cell types. Interestingly, the NF-YA subunit, binding to the promoter, is primarily a short isoform in hESCs and a long isoform in cancer cells, indicating a different activation mechanism of the CDCA8 transcription between hESCs and cancer cells. Finally, enhanced CDCA8 promoter activities by NF-Y overexpression and reduced CDCA8 transcription by NF-Y knockdown further verified that NF-Y is a positive regulator of CDCA8 transcription. Our study unearths the molecular mechanisms underlying the activation of CDCA8 expression in hESCs and cancer cells, which provides a better understanding of its biological functions.
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, the National Engineering and Research Center of Human Stem Cell, Changsha 410205, and the School of Basic Medical Science, Central South University, Changsha 410013, China.
authors.forename: Can
authors.initials: C
authors.lastname: Dai
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078.
authors.forename: Cong-Xiu
authors.initials: CX
authors.lastname: Miao
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Xu
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078.
authors.forename: Lv-Jun
authors.initials: LJ
authors.lastname: Liu
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, the School of Basic Medical Science, Central South University, Changsha 410013, China.
authors.forename: Yi-Fan
authors.initials: YF
authors.lastname: Gu
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078.
authors.forename: Di
authors.initials: D
authors.lastname: Zhou
authors.affiliation: the National Engineering and Research Center of Human Stem Cell, Changsha 410205, and.
authors.forename: Lian-Sheng
authors.initials: LS
authors.lastname: Chen
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, the National Engineering and Research Center of Human Stem Cell, Changsha 410205, and linge_csu@yahoo.com.
authors.forename: Ge
authors.initials: G
authors.lastname: Lin
authors.affiliation: From the Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, the National Engineering and Research Center of Human Stem Cell, Changsha 410205, and lugxdirector@aliyun.com.
authors.forename: Guang-Xiu
authors.initials: GX
authors.lastname: Lu
mesh_terms: D000818:Animals; D023081:CCAAT-Binding Factor / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000096:biosynthesis* / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D004622:Embryo, Mammalian / Q000196:embryology; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D064593:Heterografts; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D011401:Promoter Regions, Genetic; D015533:Transcriptional Activation
chemical_list: D023081:CCAAT-Binding Factor; C490157:CDCA8 protein, human; D018797:Cell Cycle Proteins; C492061:NFYA protein, human
keywords: CDCA8; NF-Y; cancer; differentiation; embryonic stem cell; promoter; transcription