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目的研究发生伊马替尼(IM)耐药的慢性粒细胞白血病(CML)患者 BCR/ABL 融合基因 ABL 激酶区发生点突变的情况。方法采集11例(血液学耐药7例,遗传学耐药4例)共计17份发生 IM 耐药的 CML 患者 IM 治疗前后的骨髓,采用半筑巢式扩增长片段逆转录-PCR(RT-PCR)的方法,应用分别位于 BCR 基因和 ABL 基因的引物进行2次 PCR,扩增 BCR/ABL 基因 ABL 激酶区周围863 bp 碱基,进行纯化、测序序列同源性分析。结果本研究共发现3种突变,即 G250E、E255K和 T315I。其中,血液学耐药发生突变的频率为4/7,95%可信区间为18%～90%;而遗传学耐药发生突变的频率为1/4,95%可信区间为1%～81%。所有患者发生耐药前均未发生点突变。结论发生IM 耐药的 CML 患者 BCR/ABL 基因 ABL 激酶区周围存在高频率的点突变。通过对影响与 IM 结合的突变情况进行早期检测,有利于在发生耐药前进行治疗干预,为患者提供更有效的治疗选择。
Objective To investigate the point mutations in ABL kinase domain of BCR / ABL fusion gene in patients with imatinib (IM) -resistant chronic myeloid leukemia (CML). Methods Totally 17 cases of IM-resistant CML patients were collected before and after treatment in 11 cases (hematological resistance in 7 cases and genetics in 4 cases), and the bone marrow was collected before and after IM treatment. Semi-nested amplified fragment RT-PCR -PCR) were used to amplify the 863 bp base around the ABL kinase region of BCR / ABL gene by PCR using two primers located in BCR gene and ABL gene, respectively, for purification and sequence homology analysis. Results Three mutations were found in this study, namely G250E, E255K and T315I. Among them, the hematological resistance mutation frequency was 4/7, 95% confidence interval of 18% to 90%; and genetic resistance mutation frequency of 1/4, 95% confidence interval of 1% 81%. No mutation occurred in all patients before drug resistance occurred. Conclusions There is a high frequency of point mutation around the ABL kinase region of BCR / ABL gene in IM-resistant CML patients. Early detection of mutations that affect the combination with IM will facilitate therapeutic intervention before resistance develops, providing patients with more effective treatment options.