Detection of functional PTEN lipid phosphatase protein and enzyme activity in squamous cell carcinomas of the head and neck, despite loss of heterozygosity at this locus

Br J Cancer. 2001 Jun 15;84(12):1630-4. doi: 10.1054/bjoc.2001.1848.

Abstract

The human tumour suppressor gene PTEN located at 10q23 is mutated in a variety of tumour types particularly metastatic cases and in the germline of some individuals with Cowdens cancer predisposition syndrome. We have assessed the status of PTEN and associated pathways in cell lines derived from 19 squamous cell carcinomas of the head and neck. Loss of heterozygosity is evident at, or close to the PTEN gene in 5 cases, however there were no mutations in the remaining alleles. Furthermore by Western analysis PTEN protein levels are normal in all of these SCC-HN tumours and cell lines. To assess the possibility that PTEN may be inactivated by another mechanism, we characterized lipid phosphatase levels and from a specific PIP3 biochemical assay it is clear that PTEN is functionally active in all 19 human SCCs. Our data strongly suggest the possibility that a tumour suppressor gene associated with development of SCC-HN, other than PTEN, is located in this chromosomal region. This gene does not appear to be MXI-1, which has been implicated in some other human tumour types. PTEN is an important negative regulator of PI3Kinase, of which subunit alpha is frequently amplified in SCC-HN. To examine the possibility that PI3K is upregulated by amplification in this tumour set we assessed the phosphorylation status of Akt, a downstream target of PI3K. In all cases there is no detectable increase in Akt phosphorylation. Therefore there is no detectable defect in the PI3K pathway in SCC-HN suggesting that the reason for 3q26.3 over-representation may be due to genes other than PI3K110alpha.

MeSH terms

  • Blotting, Western
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / pathology
  • Chromosomes, Human, Pair 3 / genetics*
  • DNA, Neoplasm / genetics*
  • Gene Expression Regulation, Neoplastic*
  • Genetic Predisposition to Disease
  • Head and Neck Neoplasms / genetics*
  • Head and Neck Neoplasms / pathology
  • Humans
  • Loss of Heterozygosity
  • PTEN Phosphohydrolase
  • Phosphatidate Phosphatase / analysis
  • Phosphatidate Phosphatase / metabolism*
  • Phosphoric Monoester Hydrolases / analysis
  • Phosphoric Monoester Hydrolases / biosynthesis*
  • Phosphorylation
  • Protein Kinases / biosynthesis
  • Tumor Cells, Cultured
  • Tumor Suppressor Proteins*
  • Up-Regulation

Substances

  • DNA, Neoplasm
  • Tumor Suppressor Proteins
  • Protein Kinases
  • lipid phosphate phosphatase
  • Phosphoric Monoester Hydrolases
  • Phosphatidate Phosphatase
  • PTEN Phosphohydrolase
  • PTEN protein, human