摘要目的 探讨三氧化二砷(As2O3)在体外对伊马替尼(IM)耐药bcr-abl基因突变细胞株(简称IM耐药细胞株)的生长抑制作用.方法 采用MTT比色法观察IM敏感细胞株32Dp210和包括32Dp210T3151等15种IM耐药细胞株的细胞增殖能力.选择其中5种慢性粒细胞白血病(CML)常见基因突变型细胞株,经膜联蛋白V(Annexin V)标记流式细胞术检测细胞凋亡,Westem blot检测bcr-abl融合蛋白、CRKL磷酸化及细胞凋亡相关蛋白表达的,变化.结果 32Dp210T3151等15种IM耐药细胞株具有不同程度的IM耐药性(IC50为IM敏感细胞株32Dp210的1.5～25.0倍).As2O3对IM耐药细胞株的IC50值仅为32Dp210细胞的0.18～0.34.将5种CML常见的IM耐药细胞株与32Dp210比较发现,As2O3更显著地旱剂量依赖性抑制IM耐药细胞株bcr-abl融合蛋白、磷酸化CRKL蛋白的表达,诱导细胞凋亡的作用也明显强于32Dp210.4 μmol/L As2O3可激活5种突变细胞的caspase-3、8、9蛋白表达.结论 As2O3在体外对IM耐药细胞株具有比IM敏感细胞株更显著的生长抑制和诱导凋亡的作用,细胞凋亡的产生主要与caspase-3、8、9蛋白的激活有关,提示As2O3可能成为治疗IM耐药CML患者的新选择.

abstractsObjective To explore the effect of arsenic trioxide (As2O3) on the growth inhibition of imatinib(IM)-resistant bcr-abl mutant cell lines in vitro. Methods Cell growth of one IM-sensitive cell line, 32Dp210 and 15 IM-resistant cell lines including 13151 and other 14 bcr-abl mutants were detected by MTT assay after treatment with IM and As2O3. The cell lines with 5 frequently observed mutants in CML pa-tients were analyzed for apoptosis by flow cytometry with Annexin V and PI staining as well as the expression of bcr-abl fusion protein, phosphorylated CRKL protein and apoptosis-related proteins by Western blot. Re-suits The fifty percent inhibition concentration (IC50) values of As2O3 for 15 IM-resistant cell lines were 2.6 - 5.3 fold lower than that for IM-sensitive cell line. For the 5 bcr-abl mutants frequently happened in CML patients, As2O3 significantly inhibited the expression of bcr-abl fusion protein and phosphorylated CRKL and induced apoptosis in a dose-dependent manner as compared with that for 320p210. Coincidently, the cell apeptosis was induced through caspase-3, 8 and 9 pathways. Conclusion As2O3 remarkably inhibits cell growth and induces apoptosis of IM-resistant ber-abl mutant cell lines in vitro, suggesting that it might be a potential therapeutic agent for IM-resistant CML patients.