WT1 regulates cyclin A1 expression in K562 cells

Oncol Rep. 2019 Nov;42(5):2016-2028. doi: 10.3892/or.2019.7287. Epub 2019 Aug 21.

Abstract

The restricted expression of Wilms tumor 1 (WT1) and cyclin A1 (CCNA1) in normal tissues, as opposed to their abnormal expression in leukemia demonstrates the applicability of WT1 and CCNA1 as cancer antigens for immunotherapy, and as markers for prognosis and relapse. In this study, the WT1 and CCNA1 mRNA levels were found to be elevated in bone marrow samples from pediatric acute promyelocytic leukemia (APL or AML‑M3) patients, and to be quite varied in pediatric acute lymphocytic leukemia (ALL) patients, compared to non‑leukemic bone marrow controls. Consistent with the observed upregulation of both WT1 and CCNA1 in APL, WT1 overexpression elevated the CCNA1 mRNA levels in K562 leukemia cells. Treatment with curcumin decreased the WT1 levels in K562 cells, and also decreased CCNA1 protein expression. The examination of the CCNA1 promoter identified potential canonical WT1 binding sites within the 3‑kb region upstream of the transcription start site. Chromatin immunoprecipitation and luciferase reporter assays confirmed WT1 binding and the activation of the CCNA1 promoter. Furthermore, the GC‑rich core CCNA1 promoter region provided additional non‑canonical WT1 activation sites, as revealed by promoter assays. The importance of the GC‑rich core region of the CCNA1 promoter was confirmed by treating the K562 cells with mithramycin A, which blocks the binding of zinc finger transcription factors to GC‑rich sequences. Mithramycin A subsequently suppressed both CCNA1 promoter activity and protein expression in the K562 cells. Taken together, the data from the WT1 overexpression, and curcumin and mithramycin A treatment experiments, as well as those from chromatin binding assays, along with inferences from patient RNA analyses, establish a plausible link between WT1 and CCNA1, and support the functional significance of an elevated WT1 expression in leukemia, which may also affect CCNA1 expression.

MeSH terms

  • Adolescent
  • Binding Sites
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Child
  • Child, Preschool
  • Curcumin / pharmacology
  • Cyclin A1 / chemistry
  • Cyclin A1 / genetics*
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Infant
  • K562 Cells
  • Leukemia, Promyelocytic, Acute / genetics*
  • Leukemia, Promyelocytic, Acute / metabolism
  • Male
  • Plicamycin / analogs & derivatives
  • Plicamycin / pharmacology
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / genetics*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / metabolism
  • Promoter Regions, Genetic / drug effects
  • Up-Regulation / drug effects
  • WT1 Proteins / chemistry
  • WT1 Proteins / genetics*
  • WT1 Proteins / metabolism

Substances

  • CCNA1 protein, human
  • Cyclin A1
  • WT1 Proteins
  • WT1 protein, human
  • mithramycin A
  • Curcumin
  • Plicamycin