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目的探讨MTHFR C677T和RFC1G80A基因多态性是否对儿童ALL大剂量甲氨蝶呤(HD-MTX)化疗后不良反应有影响。方法回顾性分析53例ALL患儿(2~12岁)HD-MTX化疗后的不良反应,并检测患儿MTHFR C677T和RFC1G80A基因型,探讨不良反应与基因型的相关性。采用PCR-限制性内切酶片段长度多态性技术(PCR-RFLP)扩增基因片段并酶切电泳观察分析MTHFR C677T和RFC1 G80A的多态性。患儿HD-MTX化疗后发生的不良反应按照WHO标准统一评价,采用免疫偏振荧光法(FPIA)检测HD-MTX化疗44h后MTX血清药物水平。结果53例ALL患儿中,MTHFR677CC、677CT和677TT基因型分别占28.3%、43.4%和28.3%;RFC180GG、80GA和80AA基因型分别占13.2%、58.5%和28.3%。接受HD-MTX化疗的53例ALL患儿中,总体不良反应发生率为62.3%(33/53例),而RFC180AA基因型患儿发生不良反应的可能性是RFC180GG基因型患儿的10倍(OR=10.00,95%CI:1.26~79.34,P=0.03)。在有不良反应的患儿中,RFC180A等位基因携带频率(70.5%)显著高于RFC180G(51.1%,P=0.04),携带RFC180A等位基因可增加HD-MTX化疗后不良反应的发生风险(OR=2.29,95%CI:1.02~5.10)。与其他基因型患儿比较,RFC180AA基因型者发生Ⅱ度以上肝损伤的风险加大(OR=5.6,95%CI:1.536~20.420,P=0.006),并且发生排泄延迟的概率也较其他基因型者高(OR=5.061,95%CI:1.373~18.654,P=0.028)。结论RFC1G80A基因多态性有望成为对ALL患儿HD-MTX毒性反应进行预测的有效指标之一。
Objective To investigate whether MTHFR C677T and RFC1G80A polymorphisms affect the adverse reactions of high-dose methotrexate (HD-MTX) chemotherapy in pediatric ALL patients. Methods The adverse reactions after HD-MTX chemotherapy in 53 children with ALL (range 2-12 years) were retrospectively analyzed. The genotypes of MTHFR C677T and RFC1G80A in children were detected, and the association between adverse reactions and genotypes was also explored. PCR-RFLP was used to amplify the gene fragment and analyze the polymorphisms of MTHFR C677T and RFC1 G80A by enzyme-cut electrophoresis. Adverse reactions after HD-MTX chemotherapy in children were evaluated according to the WHO standard. MTX serum drug levels were measured 44 days after HD-MTX chemotherapy by using polarized fluorescence immunoassay (FPIA). Results Among 53 children with ALL, MTHFR677CC, 677CT and 677TT genotypes accounted for 28.3%, 43.4% and 28.3%, respectively. The genotypes of RFC180GG, 80GA and 80AA accounted for 13.2%, 58.5% and 28.3% respectively. The overall incidence of adverse events in 53 ALL patients receiving HD-MTX chemotherapy was 62.3% (33/53 patients), while the incidence of adverse reactions in children with RFC180AA genotype was 10 times more likely to be children with RFC180GG genotype OR = 10.00, 95% CI: 1.26-79.34, P = 0.03). In children with adverse reactions, the frequency of RFC180A alleles (70.5%) was significantly higher than that of RFC180G (51.1%, P = 0.04), and carrying the RFC180A allele increased the risk of adverse events after HD-MTX chemotherapy OR = 2.29, 95% CI: 1.02 ~ 5.10). Compared with children with other genotypes, the risk of developing orthotopic liver cirrhosis was significantly higher in patients with RFC180AA genotype (OR = 5.6, 95% CI: 1.536-20.420, P = 0.006), and the probability of excretion delay was higher than those in other genotypes Type (OR = 5.061, 95% CI: 1.373-18.654, P = 0.028). Conclusion RFC1G80A gene polymorphism is expected to be one of the effective indicators for predicting HD-MTX toxicity in children with ALL.