A chimeric protein PTEN-L-p53 enters U251 cells to repress proliferation and invasion

Exp Cell Res. 2018 Aug 15;369(2):234-242. doi: 10.1016/j.yexcr.2018.05.023. Epub 2018 May 23.

Abstract

PTEN, a well-known tumor suppressor, dephosphorylates PIP3 and inhibits AKT activity. A translational variant of PTEN has been identified and termed PTEN-Long (PTEN-L). The additional 173 amino acids (PTEN-L leader) at the N-terminal constitute a potential signal peptide. Differing from canonical PTEN, PTEN-L is secreted into the extracellular fluid and re-enters recipient cells, playing the similar roles as PTEN in vivo and in vitro. This character confers the PTEN-L a therapeutic ability via directly protein delivering instead of traditional DNA and RNA vector options. In the present study, we employed PTEN-L leader to assemble a fusion protein, PTEN-L-p53, inosculated with the transcriptional regulator TP53, which is another powerful tumor suppressor. We overexpressed PTEN-L-p53 in HEK293T cells and detected it in both the cytoplasm and nucleus. Subsequently, we found that PTEN-L-p53 was secreted outside of the cells and detected in the culture media by immunoblotting. Furthermore, we demonstrated that PTEN-L-p53 freely entered the cells and suppressed the viability of U251cells (p53R273H, a cell line with p53 R273H-mutation). PTEN-L-p53 is composed of endogenous protein/peptide bearing low immunogenicity, and only the junction region between PTEN-L leader and p53 can act as a new immune epitope. Accordingly, this fusion protein can potentially be used as a therapeutic option for TP53-abnormality cancers.

Keywords: Cancer; PTEN-L-p53; PTEN-Long; TP53; U251 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Movement / immunology
  • Cell Movement / physiology
  • Cell Proliferation / genetics
  • Cell Proliferation / physiology
  • Drug Delivery Systems
  • HEK293 Cells
  • Humans
  • Mutation, Missense
  • Neoplasm Invasiveness / genetics
  • Neoplasm Invasiveness / immunology
  • Neoplasm Invasiveness / physiopathology
  • PTEN Phosphohydrolase / genetics
  • PTEN Phosphohydrolase / immunology
  • PTEN Phosphohydrolase / metabolism*
  • Protein Sorting Signals / genetics
  • Protein Sorting Signals / physiology
  • Protein Transport
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / immunology
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • PTEN Phosphohydrolase
  • PTEN protein, human