DHPLC analysis of the matrix metalloproteinase-1 promoter 1G/2G polymorphism that can be easily used to screen large population

J Biochem Biophys Methods. 2005 Jun 30;63(3):222-7. doi: 10.1016/j.jbbm.2005.03.007.

Abstract

Matrix metalloproteinase-1 has been shown to play an important role in all stages of cancer initiation, invasion, and metastasis. The 1G/2G single nucleotide polymorphism (SNP) at -1607 to -1608 creates an Ets binding site and elevates the rate of transcription. Moreover, the presence of the 2G allele in the MMP-1 promoter has been reported to be associated with the development and/or progression of carcinomas of the ovary, endometrium, lung, and colorectum. However, further studies on a wide variety of cancers in various sufficiently large populations will be required to verify that 2G is risk factor for cancers. A major challenge confronting such studies is the need to develop accurate, fast and inexpensive high-throughput genotyping techniques. To set up a fast and sensitive test for MMP-1 1G/2G genotyping, we analyzed 126 healthy persons by denaturing high performance liquid chromatography (DHPLC). The genotypes of MMP-1 1G/2G revealed by DHPLC analysis were further confirmed by DNA sequencing. In conclusion, DHPLC is a cost-effective, rapid, sensitive, and high-throughput technique for MMP-1 1G/2G genotyping.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Base Sequence
  • Binding Sites
  • Chromatography, High Pressure Liquid / methods*
  • Genetic Predisposition to Disease
  • Genotype
  • Humans
  • Mass Screening
  • Matrix Metalloproteinase 1 / genetics*
  • Molecular Sequence Data
  • Neoplasms / genetics*
  • Polymerase Chain Reaction
  • Polymorphism, Genetic*
  • Polymorphism, Single Nucleotide
  • Promoter Regions, Genetic*
  • Risk Factors
  • Sequence Analysis, DNA

Substances

  • Matrix Metalloproteinase 1