LNK基因在慢性髓系白血病中的变异

摘要

Objective:To compare the mutation and single nucleotide polymorphism (SNP) of LNK gene between chronic myeloid leukemia(CML) and control groups,and to explore the relationship between LNK gene variation and the occurrence of CML.Methods:A total of 36 patients with CML were selected,46 healthy persons were used as normal controls.DNA was extracted from bone marrow and peripheral blood,BCR/ABL1 fusion gene was detected by Q-PCR.The whole exon of LNK gene was amplified by PCR.The amplified sequences included the Rs3184504 (C/T) and Rs78894077 (A/C/G/T) affecting the expression of amino acids in LNK gene,and the Rs7973120 (A/T) unaffecting the expression of the amino acids.The mutations and SNP of LNK gene were analyzed by DNA sequencing.Results:Thirty-six cases of CML had BCR/ABL1 mutation,while no mutation was found in the control group.One case of CML had LNK heterozygous mutation (A300V),and the mutation rate was 2.8％,no mutation was seen in normal control group.Rs3184504:C/T allele frequency was 50％/50％ in the control group,94.4％/5.6％ in the CML group,and the C allele in CML group was significantly higher than that in the control group;CC genotype accounted for 94.4％ (P ＜0.01).Rs78894077:C/T allele in the control group was 9.8％/90.2％,in CML group was 16.7％/83.3％,the difference was not statistically significant(P ＞ 0.05);but CC genotype in CML group was very statistically significant higher than that in control group(P ＜ 0.01).Rs7973120:A/T allele frequency was 10.9％/89.1％ in the control group,25％/75％ in the CML group,the LNK A allele in CML group was very significantly higher than that in control group (P ＜ 0.01).Conclusion:CML patients have been confirmed to have LNK mutation;the SNPs of LNK are related with the development of CML,and the most CML patients carry the LNK Rs3184504 C allele and the Rs7973120 A allele.%目的:比较慢性髓系白血病(CML)患者组与对照组的LNK基因突变及单核苷酸多态性(SNP),探讨LNK基因变异与CML发生的关系.方法:选取36例CML患者和46例健康对照者.提取骨髓和外周血DNA,用Q-PCR检测BCR/ABL1融合基因,用PCR扩增LNK基因外显子全长;扩增序列中包括了LNK基因内影响氨基酸表达的Rs3184504 (C/T)和Rs 78894077 (A/C/G/T),以及对氨基酸表达无影响的Rs7973120(A/T)的3个SNP位点.测序分析LNK基因突变及单核苷酸多态性.结果:36例CML患者均有BCR/ABL1突变,对照组无突变;1例CML患者有LNK杂合子突变,位点为A300V,突变率2.8％,对照组无突变.Rs3184504:对照组C/T等位基因频率为50％/50％,CML组为94.4％/5.6％,CML组C等位基因明显高于对照组,其中CC基因型占94.4％ (P ＜0.01);Rs78894077:对照组C/T等位基因为9.8％/90.2％,CML组16.7％/83.3％,差异无统计学意义(P＞0.05),但CML组CC基因型高于对照组,差异有统计学意义(P＜0.01);Rs7973120:对照组A/T等位基因频率为10.9％/89.1％,CML组为25％/75％,CML组A等位基因高于对照组(P＜0.01).结论:CML患者中有LNK突变,LNK单核苷酸多态性与CML的发生相关,CML患者多携带有LNK Rs3184504C等位基因及Rs7973120 A等位基因.