Live cell, image-based high-throughput screen to quantitate p53 stabilization and viability in human papillomavirus positive cancer cells

Virology. 2021 Aug:560:96-109. doi: 10.1016/j.virol.2021.05.006. Epub 2021 May 22.

Abstract

Approximately 5% of cancers are caused by high-risk human papillomaviruses. Although very effective preventive vaccines will reduce this cancer burden significantly over the next several decades, they have no therapeutic effect for those already infected and remaining at risk for malignant progression of hrHPV lesions. HPV-associated cancers are dependent upon the expression of the viral E6 and E7 oncogenes. The oncogenic function of hrHPV E6 relies partially on its ability to induce p53 degradation. Since p53 is generally wildtype in hrHPV-associated cancers, p53 stabilization arrests proliferation, induces apoptosis and/or results in senescence. Here we describe a live cell, image-based high-throughput screen to identify compounds that stabilize p53 and/or affect viability in HPV-positive cancer HeLa cells. We validate the robustness and potential of this screening assay by assessing the activities of approximately 6,500 known bioactive compounds, illustrating its capability to function as a platform to identify novel therapeutics for hrHPV.

Keywords: Cervical cancer; E6 oncoprotein; E6AP; High-throughput screen; Human papillomavirus; Small molecule; p53.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Apoptosis / drug effects
  • Aurora Kinases / antagonists & inhibitors*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cyclin-Dependent Kinases / antagonists & inhibitors*
  • DNA-Binding Proteins / metabolism
  • Female
  • HeLa Cells
  • High-Throughput Screening Assays / methods*
  • Histone Deacetylase Inhibitors / pharmacology*
  • Human papillomavirus 18 / genetics
  • Humans
  • Oncogene Proteins, Viral / metabolism
  • Papillomavirus Infections / diagnosis
  • Papillomavirus Infections / diagnostic imaging
  • Papillomavirus Infections / pathology
  • Topoisomerase Inhibitors / pharmacology*
  • Tumor Suppressor Protein p53 / metabolism*
  • Uterine Cervical Neoplasms / genetics
  • Uterine Cervical Neoplasms / virology

Substances

  • DNA-Binding Proteins
  • E6 protein, Human papillomavirus type 18
  • E7 protein, Human papillomavirus type 18
  • Histone Deacetylase Inhibitors
  • Oncogene Proteins, Viral
  • TP53 protein, human
  • Topoisomerase Inhibitors
  • Tumor Suppressor Protein p53
  • Aurora Kinases
  • Cyclin-Dependent Kinases