CAJ | 학술논문

目的探讨c-Myc在骨髓基质细胞介导的急性髓细胞白血病(AML)细胞系耐药中的作用,从肿瘤微环境的角度探索AML耐药的分子机制.方法将AML细胞系U937、KG1a与造血干细胞移植健康供者的骨髓基质细胞(间充质干细胞,MSC)共培养,以细胞单独培养作为对照.流式细胞术及膜联蛋V (AnnexinV)/碘化丙啶(PI)双染色法和DAPI染色法比较两种培养条件下AML细胞对米托蒽醌诱导的凋亡的差异；Western印迹检测两种培养条件下AML细胞c-Myc蛋白的表达；在培养体系中加入c-Myc抑制剂10058-F4,检测AML细胞对米托蒽醌诱导的凋亡的变化.结果 U937、KG1a两种AML细胞系与MSC共培养后,对米托蒽醌诱导的凋亡均低于对照组(9.88％±1.53％比42.83％±2.03％,P=0.004；20.60％±2.87％比42.53％±5.29％,P=0.030),共培养促进了AML细胞对化疗药物的耐药性.AML细胞系与MSC共培养后,Western印迹检测c-Myc蛋白的表达明显高于对照组.c-Myc抑制剂10058-F4可诱导AML细胞的凋亡；10058-F4加入共培养体系后,KG1a细胞系对米托蒽醌诱导的凋亡率从23.87％±1.55％显著升高到57.23％±3.88％(P=0.009),在U937细胞共培养体系中同样观察到细胞凋亡率从16.07％±2.11％显著提高到53.47％±4.08％ (P =0.004),从而克服耐药.结论 AML细胞与MSC共培养可导致c-Myc蛋白表达的上调,从而介导AML细胞对化疗药物的耐药；针对c-Myc的靶向治疗将为AML的治疗提供新思路.
목적 탐토c-Myc재골수기질세포개도적급성수세포백혈병(AML)세포계내약중적작용,종종류미배경적각도탐색AML내약적분자궤제.방법 장AML세포계U937、KG1a여조혈간세포이식건강공자적골수기질세포(간충질간세포,MSC)공배양,이세포단독배양작위대조.류식세포술급막련단V (AnnexinV)/전화병정(PI)쌍염색법화DAPI염색법비교량충배양조건하AML세포대미탁은곤유도적조망적차이；Western인적검측량충배양조건하AML세포c-Myc단백적표체；재배양체계중가입c-Myc억제제10058-F4,검측AML세포대미탁은곤유도적조망적변화.결과 U937、KG1a량충AML세포계여MSC공배양후,대미탁은곤유도적조망균저우대조조(9.88％±1.53％비42.83％±2.03％,P=0.004；20.60％±2.87％비42.53％±5.29％,P=0.030),공배양촉진료AML세포대화료약물적내약성.AML세포계여MSC공배양후,Western인적검측c-Myc단백적표체명현고우대조조.c-Myc억제제10058-F4가유도AML세포적조망；10058-F4가입공배양체계후,KG1a세포계대미탁은곤유도적조망솔종23.87％±1.55％현저승고도57.23％±3.88％(P=0.009),재U937세포공배양체계중동양관찰도세포조망솔종16.07％±2.11％현저제고도53.47％±4.08％ (P =0.004),종이극복내약.결론 AML세포여MSC공배양가도치c-Myc단백표체적상조,종이개도AML세포대화료약물적내약；침대c-Myc적파향치료장위AML적치료제공신사로.
Objective To explore the role of c-Myc in mesenchymal stromal cell-mediated drug resistance and elucidate the molecular mechanism of acute myeloid leukemia (AML) from the version of tumor microenvironment.Methods AML cell lines U937 and KG1a were co-cultured with mesenchymal stromal cells (MSC) from bone marrow of healthy donors between January to March 2012.The AML cell lines plated alone was cultured as controls.Apoptosis induced by mitoxantrone was measured by flow cytometry and Annexin V/PI double and 4'-6-diamidino-2-phenylindole (DAPI) staining,And c-Myc protein was detected by Western blot under both culturing conditions.After a pre-treatment of c-Myc inhibitor 10058-F4,the apoptosis of AML cell was also evaluated.Results Apoptosis of AML cells (U937 and KG1a) significantly decreased during co-culturing with MSC (9.88％ ± 1.53％ vs 42.83％ ± 2.03％,P =0.004;20.60％ ±2.87％ vs 42.53％ ±5.29％,P =0.030).Drug resistance was implicated.The coculturing of AML cells with MSC significantly induced an up-regulation of c-Myc.The inhibition of c-Myc with 10058-F4 could induce apoptosis of AML cells.After an addition of 10058-F4 into the co-culture system,the apoptotic rate of KG1a cells significantly increased from 23.87％ ± 1.55％ to 57.23％ ± 3.88％ (P =0.009).Similarly the apoptotic rates spiked from 16.07％ ± 2.11％ to 53.47％ ± 4.08％ in U937 cells (P =0.004) to overcome the stromal cell-mediated drug resistance.Conclusions The co-culturing of AML cells and MSC induces an up-regulation of c-Myc protein so as to cause the emergence of chemoresistance.Therefore targeting c-Myc orotein may provide a novel theraoeutic strategy of AML.