PCR-SSCP-DNA sequencing method in detecting PTEN gene mutation and its significance in human gastric cancer

World J Gastroenterol. 2008 Jun 28;14(24):3804-11. doi: 10.3748/wjg.14.3804.

Abstract

Aim: To discuss the possible effect of PTEN gene mutations on occurrence and development of gastric cancer.

Methods: Fifty-three gastric cancer specimens were selected to probe PTEN gene mutations in genome of gastric cancer and paracancerous tissues using PCR-SSCP-DNA sequencing method based on microdissection and to observe the protein expression by immunohistochemistry technique.

Results: PCR-SSCP-DNA sequencing indicated that 4 kinds of mutation sites were found in 5 of 53 gastric cancer specimens. One kind of mutation was found in exons. AA-TCC mutation was located at 40 bp upstream of 3' lateral exon 7 (115946 AA-TCC). Such mutations led to terminator formation in the 297th codon of the PTEN gene. The other 3 kinds of mutation were found in introns, including a G-C point mutation at 91 bp upstream of 5' lateral exon 5(90896 G-C), a T-G point mutation at 24 bp upstream of 5' lateral exon 5 (90963 T-G), and a single base A mutation at 7 bp upstream of 5' lateral exon 5 (90980 A del). The PTEN protein expression in gastric cancer and paracancerous tissues detected using immunohistochemistry technique indicated that the total positive rate of PTEN protein expression was 66% in gastric cancer tissue, which was significantly lower than that (100%) in paracancerous tissues (P < 0.005).

Conclusion: PTEN gene mutation and expression may play an important role in the occurrence and development of gastric cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Base Sequence
  • Exons / genetics
  • Female
  • Humans
  • Male
  • Middle Aged
  • Mutation / genetics*
  • PTEN Phosphohydrolase / genetics*
  • Polymerase Chain Reaction
  • Polymorphism, Single-Stranded Conformational
  • Sequence Analysis, DNA
  • Stomach Neoplasms / genetics*

Substances

  • PTEN Phosphohydrolase
  • PTEN protein, human