摘要
目的应用简易的HPLC方法检测血红细胞中巯嘌呤甲基转移酶(TPMT)的活性,并研究健康汉族成年人群和成人急性淋巴细胞白血病(ALL)患者TPMT活性分布。方法以非放射性S-腺苷-L-甲硫氨酸(SAM)为甲基供体,完成6-巯基嘌呤(6-MP)向6-甲基巯基嘌呤(6-MeMP)的甲基化转变。6-MeMP经乙酸苯汞混和物抽提,采用高效液相色谱法分析。TPMT的活性用37℃时每小时每毫升红细胞生成多少纳摩尔的6-MeMP来表示。结果健康汉族成人和成人ALL患者TPMT活性均呈正态分布,男性和女性相比,TPMT活性没有显著性差异。健康汉族成人与成人ALL患者服用6-MP前TPMT活性相比没有显著性差异。而TPMT*3C杂合子TPMT平均活性低于野生型TPMT基因(TPMT*1)纯合子,差异有显著性。结论TPMT活性在健康汉族成人和成人ALL患者均呈正态分布,TPMT基因型和表型存在相关性,TPMT*3C杂合子平均酶活性低于野生型TPMT基因(TPMT*1)纯合子。在开始6-MP等药物维持治疗前检测TPMT活性对成人ALL患者有益。
Aim To employ a simple HPLC assay to measure thiopurine S-methyltransferase（TPMT） activity.The assay was used to study the distribution of TPMT activity in an adult acute lymphoblastic leukemia population.Methods The assay is based on the conversion of 6-mercatopurine（6-MP） to 6-methylmercatopurine（methyl-MP）,using S-adenosyl-L-methionine （SAM） as the methyl donor.Methyl-MP was extracted into toluene as a phenyl-mercury adduct.Methyl-MP was quantitated by reversed-phase high-performance liquid chromatography（HPLC）.One unit of enzyme activity represented the formation of 1 nmol of methyl-MP per hour of incubation,per milliliter of packed red cells at 37℃.Result The frequency distribution histogram,the probit plot and the Kolmogorov-Smirnov test supported a normal distribution of TPMT activity in this Han Chinese population sample of healthy and adult ALL.There was not a significant difference in TPMT activity between men and women within each group involved in this study.There was not a significant difference in TPMT activity between Han Chinese healthy adults and ALL at diagnosis.However,TPMT activity of those subjects with TPMT＊1/TPMT＊3C was lower than the mean value of those subjects with TPMT＊1/ TPMT＊1 in both Han Chinese adults and ALL.Conclusions A normal distribution was obtained of TPMT activity in this Han Chinese population sample of healthy and adult ALL.TPMT activity of those subjects with TPMT＊1/TPMT＊3C was lower than the mean value of those subjects with TPMT＊1/ TPMT＊1 in both Han Chinese adults and ALL.Prospective identification of TPMT activity may have clinical benefit to Chinese adult ALL patients treated with thiopurine drugs.
出处
《安徽医药》
CAS
2008年第9期785-787,共3页
Anhui Medical and Pharmaceutical Journal
基金
安徽省教育厅自然科学基金(编号2003kj192和2005kj338)
