title: 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.
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.
authors.affiliation: Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, 400038, Chongqing, PR China. a65424208@online.cq.cn
authors.forename: Ze-Jun
authors.initials: ZJ
authors.lastname: Liu
authors.forename: Wei-Cong
authors.initials: WC
authors.lastname: Peng
authors.forename: Xin
authors.initials: X
authors.lastname: Yang
authors.forename: Jun-Fu
authors.initials: JF
authors.lastname: Huang
authors.forename: Xiao-Bing
authors.initials: XB
authors.lastname: Zhang
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Masato
authors.initials: M
authors.lastname: Maekawa
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers; D018076:DNA, Complementary; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D000080864:Lactate Dehydrogenase 5; D018807:Polymorphism, Single-Stranded Conformational; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D007770:L-Lactate Dehydrogenase; D000080864:Lactate Dehydrogenase 5; C409667:lactate dehydrogenase 1