ALPK2 is crucial for luminal apoptosis and DNA repair-related gene expression in a three-dimensional colonic-crypt model

Anticancer Res. 2012 Jun;32(6):2301-8.

Abstract

Background: Oncogenic KRAS signaling is dysregulated in a three-dimensional (3D)-specific manner in human colorectal cancer (CRC) HCT116 cells. However, the identity of the crucial genes which are down-regulated through oncogenic KRAS in 3D cultures remains unclear.

Materials and methods: We established a specific anti-alpha-kinase 2 (ALPK2) antibody and addressed the ALPK2 function in HKe3 cells, which are HCT116 cells with a disruption in oncogenic KRAS, in a 3D colonic-crypt model.

Results: In HKe3 cells grown in 3D culture, ALPK2 siRNA inhibited luminal apoptosis and reduced the expression of cleaved caspase-3. Furthermore, ALPK2 siRNA reduced the expression of DNA repair genes. Reduced expression of ALPK2 mRNA was found to be correlated with clinical colorectal adenomas in a public dataset of gene expression analyses.

Conclusion: ALPK2, down-regulated by oncogenic KRAS, is crucial for luminal apoptosis and expression of DNA repair-related genes, possibly in the transition of normal colonic crypt to adenoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics*
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Transformation, Neoplastic / genetics
  • Cell Transformation, Neoplastic / pathology
  • Colorectal Neoplasms / enzymology*
  • Colorectal Neoplasms / genetics
  • DNA Repair / genetics*
  • Fluorescent Antibody Technique
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • HCT116 Cells
  • Humans
  • Immunoprecipitation
  • Microscopy, Confocal
  • Oligonucleotide Array Sequence Analysis
  • Phosphotransferases / genetics
  • Phosphotransferases / metabolism*
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins p21(ras)
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection
  • ras Proteins / genetics

Substances

  • KRAS protein, human
  • Proto-Oncogene Proteins
  • RNA, Small Interfering
  • ALPK2 protein, human
  • Phosphotransferases
  • Protein Kinases
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins