T85C polymorphisms of the dihydropyrimidine dehydrogenase gene detected in gastric cancer tissues by high-resolution melting curve analysis

Turk J Gastroenterol. 2012;23(6):652-7. doi: 10.4318/tjg.2012.0469.

Abstract

Background/aims: Dihydropyrimidine dehydrogenase is a key enzyme acting on the metabolic pathway of medications for gastric cancer. High-resolution melting curve technology, which was developed recently, can distinguish the wild-type dihydropyrimidine dehydrogenase gene from multiple polymorphisms by fluorescent quantitative polymerase chain reaction products in a direct and effective manner.

Materials and methods: T85C polymorphisms of dihydropyrimidine dehydrogenase in the peripheral blood of 112 Chinese gastric cancer patients were detected by real-time polymerase chain reaction combined with high-resolution melting curve technology. Primer design, along with the reaction system and conditions, was optimized based on the GenBank sequence.

Results: Seventy nine cases of wild-type (TT, [70.5%]), 29 cases of heterozygous (TC, [25.9%]), and 4 cases of homozygous mutant (CC, [3.6%]) were observed. The result was completely consistent with the results of the sequencing.

Conclusions: Real-time polymerase chain reaction combined with high-resolution melting curve technology is a rapid, simple, reliable, direct-viewing, and convenient method for the detection and screening of polymorphisms.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Asian People / genetics
  • Dihydrouracil Dehydrogenase (NADP) / genetics*
  • Female
  • Genetic Predisposition to Disease / genetics
  • Genetic Testing / methods*
  • Genetic Testing / standards*
  • Heterozygote
  • Homozygote
  • Humans
  • Male
  • Middle Aged
  • Polymorphism, Single Nucleotide
  • Real-Time Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / standards*
  • Reproducibility of Results
  • Stomach Neoplasms / genetics*

Substances

  • Dihydrouracil Dehydrogenase (NADP)