c-Myc抑制剂10058-F4对伊马替尼耐药细胞的增殖抑制作用  

Inhibition of c-Myc by 10058-F4 overcomes imatinib resistance in chronic myeloid leukemia cells

作　　者：龙梓洁[1,2] 方志刚[1,2] 潘晓娜[1,2] 范蕊芳[1,2] 林东军[1,2] 

机构地区：[1]中山大学附属第三医院血液科 [2]中山大学血液病研究所,广东广州510630

出　　处：《中国病理生理杂志》2014年第9期1590-1594,共5页Chinese Journal of Pathophysiology

基　　金：广州市科技计划珠江科技新星专项(No.2012J2200077)

摘　　要：目的:探讨c-Myc小分子抑制剂10058-F4对慢性粒细胞白血病细胞K562及伊马替尼耐药的K562/G细胞的影响,为克服耐药提供新的治疗策略。方法:Western blotting测定细胞中c-Myc蛋白表达水平,PI染色检测细胞周期。MTT法及克隆形成实验测定K562及K562/G细胞的增殖情况,Annexin V-PI染色检测细胞凋亡。结果:MTT结果显示,耐药细胞K562/G与K562细胞相比,对伊马替尼的敏感性明显降低;Western blotting检测结果表明耐药细胞具有c-Myc蛋白高表达的特点。进一步MTT结果显示,与对照组相比,c-Myc小分子抑制剂10058-F4可抑制K562及K562/G细胞的增殖,并呈剂量及时间依赖性。重要的是,K562/G细胞比K562细胞对10058-F4敏感性更高。细胞周期检测显示,10058-F4可使K562及K562/G细胞周期阻滞于G0/G1期,表明抑制cMyc导致的增殖抑制与细胞周期阻滞相关。Annexin V-PI染色结果显示10058-F4可促进K562及K562/G细胞发生凋亡。克隆形成实验表明K562/G耐药细胞的克隆形成数目明显高于K562细胞。同时,与对照组相比,10058-F4可抑制细胞的克隆形成能力,并增强伊马替尼的毒性作用。结论:耐药细胞K562/G具有c-Myc高表达的细胞遗传学特点;10058-F4可抑制K562及K562/G细胞增殖,促进凋亡,并抑制细胞的克隆形成能力;c-Myc抑制剂10058-F4可逆转c-Myc高表达引起的耐药。AIM： To investigate the effect of c-Myc inhibitor 10058-F4 on human chronic myeloid leukemia（ CML） K562 cells and imatinib-resistant K562/G cells. METHODS： The protein expression of c-Myc was detected by Western blotting. Cell proliferation was evaluated by MTT assay and colony formation assay. PI staining was used to determine the cell cycle distribution. Annexin V-PI staining was applied for apoptosis detection. RESULTS： Imatinib-resistant K562 / G cells displayed lower sensitivity to imatinib than K562 cells with high expression of c-Myc. Treatment with specific c-Myc inhibitor 10058-F4 inhibited the cell proliferation in a dose- and time-dependent manner,and K562 /G displayed more sensitivity to 10058-F4 than K562 cells. 10058-F4 also induced cell cycle arrest in G0/G1 phase and induced apoptotic cell death in the 2 cells. Importantly,10058-F4 suppressed the colony formation ability in K562 and K562 /G cells.CONCLUSION： c-Myc is a novel target to overcome imatinib-induced drug resistance,and c-Myc inhibitor provides a new approach in CML therapy.

关 键 词：C-MYC 10058-F4 伊马替尼耐药 慢性粒细胞白血病 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]