ZSTK474 targeting PIK3R3 inhibits the Wilms' tumor through G0 / G1 phase arrest

PLoS One. 2024 Oct 28;19(10):e0312178. doi: 10.1371/journal.pone.0312178. eCollection 2024.

Abstract

Purpose: Wilms' tumor (WT), also known as nephroblastoma, is the predominant form of primary malignant renal cancer. The unfavorable prognoses linked to anaplastic nephroblastoma and recurrent nephroblastoma emphasize the crucial requirement for the exploration of innovative treatment modalities for WT.

Methods: Our study conducted one-way Cox regression and Kaplan-Meier analyses using TARGET-WT nephroblastoma data to identify differentially expressed genes in nephroblastoma and evaluate their prognostic relevance. Utilizing the Connectivity Map database, ZSTK474 emerged as a viable therapeutic option for WT. The effect of ZSTK474 on WT and related underlying mechanisms were further investigated through in vitro and in vivo investigations.

Results: The in vivo experiment results indicated that ZSTK474 effectively inhibited subcutaneous tumor growth in WT mice. CCK-8 assays revealed two nephroblastoma cell lines exhibited half-inhibitory concentrations of 2μM and 2.51μM for ZSTK474, respectively. ZSTK474 was shown to inhibit the migration and invasion capabilities of WT cells in both Transwell and wound healing assays. Flow cytometry apoptosis and TUNEL assays demonstrated that ZSTK474 induced apoptosis in WT cells. Cell cycle analysis revealed that ZSTK474 led to the induction of G0/G1 phase arrest. Sequencing of ZSTK474-treated WiT49 cells suggested that the impact of ZSTK474 on WT might be mediated by the PI3K/Akt pathway, specifically by inhibiting PIK3R3. Knock-down of PIK3R3 confirmed that ZSTK474 downregulated PIK3R3, reducing Akt phosphorylation, cyclin D and CDK4 levels and elevating P21 expression in nephroblastoma cells. However, current research has limitations, including a lack of understanding of the long-term effects and potential resistance mechanisms of new therapies.

Conclusion: This research provides insight into the potential of ZSTK474 and other PI3K inhibitors for treating nephroblastoma.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Female
  • G1 Phase Cell Cycle Checkpoints / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Kidney Neoplasms* / drug therapy
  • Kidney Neoplasms* / genetics
  • Kidney Neoplasms* / metabolism
  • Kidney Neoplasms* / pathology
  • Mice
  • Mice, Nude
  • Phosphatidylinositol 3-Kinases* / metabolism
  • Resting Phase, Cell Cycle / drug effects
  • Triazines / pharmacology
  • Wilms Tumor* / drug therapy
  • Wilms Tumor* / genetics
  • Wilms Tumor* / metabolism
  • Wilms Tumor* / pathology
  • Xenograft Model Antitumor Assays

Substances

  • Phosphatidylinositol 3-Kinases
  • PIK3R3 protein, human
  • Triazines

Grants and funding

This study was supported by the Chengdu Women and Children's Hospital Talent Plan (No.YC2022017). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.