The Mthfr Gene Polymorphism And Clinical Significance Of Acute Lymphoblastic Leukemia In Patients With Methotrexate Chemotherapy

The objective of this study is to explore the relationship between methylenetetrahydrofolate reductase(MTHFR) polymorphisms and the methotrexate(MTX) concentration, the adverse effects among acute lymphoblastic leukenia(ALL) patients treated by high dose MTX.The first party is the determination of methotrexate in human plasma by RP-HPLC. The plasma sample was treated with 6% perchloric acid for sedimention and high speed centrifugation, and then sparated on a SinoChrom ODS-BP C18 (4.6×200mm,5μm) column, the mobile phase was consisted of acetoniting-0.16 mol·L-1 CH3COONa solution (12:88) at 1 mL·min-1, the detecting wavelength was set at 303nm and the column temperature was 35℃. Good linearity was found within 0.05～100μg·mL-1, r=0.9994. The recovery was higher than 75%, The intr-day and inter-day RSD<5%. The method is proved to be simple, fast, sensitive and reliable. It could be used for the determination of methotrexate plasma concentration and for pharmacokinetic study.The second party is the determination of methotrexate and the observation of the adverse effects on acute lymphoblastic leukenia(ALL). Twenty one patients with ALL in complete remission were studied on the adverse effect. Blood concentration of ((Methylenetetrahydrofolate reductase, MTX) was monitorred by high performance liquid chromatography in Oh,24h,48h,72h after the administration. MTX has a large individual variation in disposition, Gastrointestinal reaction oral mucosa, hepatic damage, renal damage and depression of bone marrow were noticed. It's necessary to determine blood concentration of MTX.The third party is genotype analysis of MTHFR gene for ALL patients.Twenty-one patients with a diagnosis of ALL were treated with HDMTX. Genomic deoxyribonucleic acid(DNA) was extracted from their peripheral blood and genotype analysis of MTHFR gene was done by polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP) techniques to investigate the relationship between the risk genotypes and HDMTX toxicity in ALL patients. MTHFR gene polymorphisms are associated with the toxicity of HDMTX in ALL. Genotype of MTHFR 677TT and 1298AA indicate most severe toxical response.