Molecular analysis of Tripterygium hypoglaucum (level) Hutch-induced mutations at the HPRT locus in human promyelocytic leukemia cells by multiplex polymerase chain reaction.
The genotoxicity and cytotoxicity of a Chinese medicinal herb, Tripterygium hypoglaucum (level) Hutch (THH), was investigated in human promyelocytic leukemia (HL-60) cells using the hypoxanthine-guanine phosphoribosyltransferase mutation assay. THH showed clear cytotoxicity and mutagenicity in HL-60 cells at concentrations between 6.7 and 20.0 mg/ml. When the mutants were characterized by techniques based on multiplex PCR, 46.6% of induced mutants were found to have deletions, whereas only 7.7% of spontaneous mutants showed deletions. The rest were not characterized, but were assumed to be mainly point mutations. Mapping of all intragenic deletion breakpoints showed a random distribution of breakpoints in nine exons. Deletion of exon 1 appeared as the only whole gene deletion, while deletions of exon 7/8 and 9 often occurred concomitantly (71.4%). It is concluded that THH is mutagenic in HL-60 cells, predominantly inducing deletions. Since this herb is widely used as a traditional medicine, its genotoxicity should be assessed in vivo in treated humans.
Liu
Sheng Xue
SX
Department of Hygiene Toxicology, College of Preventive Medicine, Third Military Medical University, Chongqing 400038, People's Republic of China.
Cao
Jia
J
An
Hui
H
D004273:DNA, Neoplasm / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005091:Exons / Q000235:genetics; D018922:HL-60 Cells / Q000737:chemistry / Q000187:drug effects*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology*; D016296:Mutagenesis; D009152:Mutagenicity Tests; D010936:Plant Extracts / Q000633:toxicity*; D018517:Plant Roots / Q000737:chemistry; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D017384:Sequence Deletion; D029757:Tripterygium / Q000737:chemistry*
D004273:DNA, Neoplasm; D010936:Plant Extracts; D007041:Hypoxanthine Phosphoribosyltransferase