Molecular analysis of THH-induced mutations at HPRT locus in human promyelocytic leukemia cells with multiplex polymerase chain reaction  

作　　者：刘胜学 曹佳 安辉 

出　　处：《Journal of Medical Colleges of PLA(China)》2002年第2期105-111,共7页中国人民解放军军医大学学报（英文版）

基　　金：Supported by National Natural Science Foundation (No. 39970650) and Chongqing Public Health Bureau Foundation (2000).

摘　　要：Objective: To study the genotoxicity and antitumor activity of a Chinese medicinal herb, Tripterygium Hypoglaucum (Level) Hutch (THH). Methods: The genotoxicity and antitumor activity of TH-H were investigated in human promyelocytic leukemia cells on the mutation of hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene by using single cell clone culture, two-way screening counting, multiplex PCR amplification and gel electrophoresis. Results: The results showed that different mutant spectra existed between the spontaneous mutation and induced mutation by THH. Only 7. 7% (1/13) of spontaneous mutants showed deletion mutations, whereas the induced mutants included 46.6% (27/58) deletions. Mapping of all intragenic deletion breakpoints showed a random distribution in all 9 exons, but toward the 3'-end of the HPRT gene. Deletion of exon 1 only appeared when whole gene was deleted. Deletions of exon 7/8 and 9 often showed linkage deletions (71.4%). Conclusion: THH can induce the mutation, mainly deletions, of HPRT gene in human promyelocytic leukemia cells.Objective: To study the genotoxicity and antitumor activity of a Chinese medicinal herb, Triptery-gium Hypoglaucum (Level) Hutch (THH). Methods: The genotoxicity and antitumor activity of THH were investigated in human promyelocytic leukemia cells on the mutation of hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene by using single cell clone culture, two-way screening counting, multiplex PCR amplification and gel electrophoresis. Results: The results showed that different mutant spectra existed between the spontaneous mutation and induced mutation by THH. Only 7. 7% (1/13) of spontaneous mutants showed deletion mutations, whereas the induced mutants included 46. 6% (27/58) deletions. Mapping of all intra-genic deletion breakpoints showed a random distribution in all 9 exons, but toward the 3'-end of the HPRT gene. Deletion of exon 1 only appeared when whole gene was deleted. Deletions of exon 7/8 and 9 often showed linkage deletions (71. 4%). Conclusion: THH can induce the mutation, mainly deletions, of HPRT gene in human promyelocytic leukemia cells.

关 键 词：HPRT gene Tripterygium Hypoglaucum Hutch MUTAGENESIS HL-60 cells multiplex PCR 

分 类 号：R733.7[医药卫生—肿瘤]