Relative mRNA expression of the lactate dehydrogenase A and B subunits as determined by simultaneous amplification and single strand conformation polymorphism. Relation with subunit enzyme activity

J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Aug 15;793(2):405-12. doi: 10.1016/s1570-0232(03)00358-1.

Abstract

To explore if it is correlated in human tumor cells that the expression of LDH homologous gene and LDH isoenzymes, we used RT-PCR-SSCP technique to measure the relative expression of genes with homologous sequences. The combination of PCR using common primers designed in the highly conserved regions and single-strand conformation polymorphism analysis of the products is used for quantitative determination of the proportions of LDH-A mRNA in human cancer cell lines. The proportion is compared with that of the activities of isoenzymes. The results indicated that the enzyme activity of LDH-A was consistent with mRNA levels in the human tumor cell. The present procedure using a single pair of primers for two fragments can overcome disadvantages in quantitative analysis using multiplex PCR. Template concentrations and PCR cycles did not affect the proportions of LDH-A and LDH-B in the product.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cell Line, Tumor
  • DNA Primers
  • DNA, Complementary
  • Humans
  • Isoenzymes / genetics*
  • L-Lactate Dehydrogenase / genetics*
  • Lactate Dehydrogenase 5
  • Polymorphism, Single-Stranded Conformational*
  • RNA, Messenger / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA Primers
  • DNA, Complementary
  • Isoenzymes
  • RNA, Messenger
  • L-Lactate Dehydrogenase
  • Lactate Dehydrogenase 5
  • lactate dehydrogenase 1