Induction of resistance to the Abelson inhibitor ST1571 in human leukemic cells through gene amplification
Induction of resistance to the Abelson inhibitor ST1571 in human leukemic cells through gene amplification
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DOI：
10.1182/blood.v95.5.1758.005a41_1758_1766
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发表时间：
2000-03-01
期刊：
影响因子：
20.3
通讯作者：
Gambacorti-Passerini, C
中科院分区：
文献类型：
--
作者：
le Coutre, P;Tassi, E;Gambacorti-Passerini, C
2-苯氨基嘧啶衍生物 STI571 已被证明可以选择性抑制致癌 bcr/abl 融合蛋白的酪氨酸激酶结构域。迄今为止，该抑制剂的活性已在来自白血病患者的 bcr/abl 表达细胞的体外和接种 bcr/abl 阳性细胞的裸鼠体内得到证明，然而，尚无关于白血病细胞是否会对 bcr/abl 抑制产生耐药性的信息，将人 bcr/abl 表达细胞系 LAMA84 与浓度不断增加的 STI571 一起培养，培养约 6 个月后，获得了新的细胞系，并将其命名为LAMA84R，这种新选择的细胞系对STI571（1.0μM）的IC50比亲代敏感细胞系的IC50（0.1μM）高10倍。 STI571 治疗显示增加了 LAMA84 中的早期和晚期凋亡分数，但在 LAMA84R 中没有增加。LAMA84 中细胞凋亡的诱导与 caspase 3 样活性的激活有关，而这种活性在耐药 LAMA84R 细胞系中不会出现。与 LAMA84 细胞相比，LAMA84R 细胞的 bcr/abl 蛋白和 mRNA 水平升高。使用 LAMA84R 细胞中的 BCR 和 ABL 特异性探针进行 FISH 分析，显示存在包含大约 13 至 14 个 BCR/ABL 基因拷贝的标记染色体。因此，通过基因扩增介导的 Bcr/Abl 蛋白的过度表达与人类白血病细胞对 STI571 的体外抗性相关，并且可能决定了该抗性，(C) 2000，美国血液学会。
The 2-phenylaminopyrimidine derivative STI571 has been shown to selectively inhibit the tyrosine kinase domain of the oncogenic bcr/abl fusion protein. The activity of this inhibitor has been demonstrated so far both in vitro with bcr/abl expressing cells derived from leukemic patients, and in vivo on nude mice inoculated with bcr/abl positive cells, Yet, no information is available on whether leukemic cells can develop resistance to bcr/abl inhibition, The human bcr/abl expressing cell line LAMA84 was cultured with increasing concentrations of STI571, After approximately 6 months of culture, a new cell line was obtained and named LAMA84R, This newly selected cell line showed an IC50 for the STI571 (1.0 mu M) 10-fold higher than the IC50 (0.1 mu M) of the parental sensitive cell line. Treatment with STI571 was shown to increase both the early and late apoptotic fraction in LAMA84 but not in LAMA84R, The induction of apoptosis in LAMA84 was associated with the activation of caspase 3-like activity, which did not develop in the resistant LAMA84R cell line. LAMA84R cells showed increased levels of bcr/abl protein and mRNA when compared to LAMA84 cells. FISH analysis with BCR- and ABL-specific probes in LAMA84R cells revealed the presence of a marker chromosome containing approximately 13 to 14 copies of the BCR/ABL gene. Thus, overexpression of the Bcr/Abl protein mediated through gene amplification is associated with and probably determines resistance of human leukemic cells to STI571 in vitro, (C) 2000 by The American Society of Hematology.