Medirex a.s., Department of Clinical Genetics, Galvaniho, Slovak Republic.
Neoplasma. 2011;58(6):548-53. doi: 10.4149/neo_2011_06_548.
The presence of BCR-ABL oncogene mutations in patients with chronic myeloid leukemia (CML) may be responsible for the failure of tyrosine kinase inhibitor (TKI) treatment. The aim of the study was to evaluate the frequency of BCR-ABL gene mutations in patients with CML treated with tyrosine kinase inhibitors. Our lab received 64 samples (34 women, 30 men) from patients with CML who failed or had suboptimal response to TKI treatment. The mutation analysis was performed in 61 patients with CML, 3 patients could not be tested because of inadequate RNA quality. An 866 base pair fragment containing the ABL kinase domain was amplified in a seminested RT (reverse transcriptase)-PCR and then sequenced using Applied Biosystems BigDye Terminator chemistry with two pairs of primers. We analyzed 61 patients with CML, 11 mutations were detected in 13 (21%) patients and SNP (single nucleotide polymorphism) in 6 patients (10%). In addition to 9 point mutations (G250E / F317L, F359V, L387M, Y253H, M388L, M244V, T315I, D276G), 35 bp insertion between exons 8 and 9 and deletion exon 7 were detected. Our results demonstrate that direct sequencing is suitable for routine clinical monitoring patients with CML and may be useful for optimizing therapy.
慢性髓性白血病（CML）患者存在 BCR-ABL 癌基因突变可能是酪氨酸激酶抑制剂（TKI）治疗失败的原因。本研究旨在评估接受酪氨酸激酶抑制剂治疗的 CML 患者中 BCR-ABL 基因突变的频率。我们实验室收到了 64 份来自 CML 患者的样本（34 名女性，30 名男性），这些患者对 TKI 治疗失败或反应不佳。对 61 例 CML 患者进行了突变分析，由于 RNA 质量不足，3 例患者无法进行检测。用Applied Biosystems BigDye Terminator 化学物和两对引物在半巢式 RT（逆转录酶）-PCR 中扩增包含 ABL 激酶结构域的 866 个碱基对片段，然后进行测序。我们分析了 61 例 CML 患者，在 13 例（21%）患者中检测到 11 种突变和 6 例（10%）患者中的 SNP（单核苷酸多态性）。除了 9 种点突变（G250E/F317L、F359V、L387M、Y253H、M388L、M244V、T315I、D276G）外，还检测到 8 号和 9 号外显子之间的 35 个碱基插入和 7 号外显子缺失。我们的结果表明，直接测序适用于 CML 患者的常规临床监测，可能有助于优化治疗。
