摘要
目的 探讨急性早幼粒细胞性白血病的特异融合蛋白PML RARα和PLZF RARα在三氧化二砷 (As2 O3 )效应中的可能作用。方法 应用逆病毒载体转染技术建立稳定表达PML RARα(KPML)、PLZF RARα(ΚPLZF)和空载体 (KV)的K5 6 2细胞克隆。通过活细胞计数、形态学观察、流式细胞仪检测细胞DNA含量和分化抗原。应用免疫荧光分析PML RARα蛋白的亚细胞分布。结果 1 0μmol/LAs2 O3 不诱导KV 细胞凋亡和分化 ,但明显抑制其生长。在KPML和KPLZF细胞 ,As2 O3 也产生相似的生长抑制效应 ,但其效应强度明显增加。经 1 0 μmol/LAs2 O3 处理 3d时 ,KV、KPML和KPLZF的生长抑制率分别为 32 %± 3%、5 7%± 4%和 5 4%± 6 %。 1 0 μmol/L的全反式维甲酸也显示对KV 细胞克隆的生长抑制 ,但其效应明显低于As2 O3 。同时 ,全反式维甲酸的生长抑制效应在KPML的表现较KV 明显(P <0 .0 5 ) ,但在KV 和KPLZF之间差异无显著意义。此外 ,在 1 0 μmol/LAs2 O3 作用 2d时 ,KV 和KPML细胞内PML/PML RARα蛋白颗粒明显减少甚至消失。结论 PML RARα和PLZF RARα蛋白明显加强As2 O3 对K5 6 2细胞的生长抑制效应。
Objective To illustrate the possible roles of acute promyelocytic leukemia specific chimeric proteins PML RARα and PLZF RARα in the effects of arsenic trioxide (As 2O 3). Methods K562 sublines stably expressing PML RARα (K PML ) and PLZF RARα(K PLZF )were established by retrovirus transfection with K V transfected empty vectors as controls. Effects of As 2O 3 and all trans retinoic acid (ATRA) on these sublines were analyzed through cell count, morphology, measurement of cellular DNA contents and differentiation antigens on flow cytometry. Subcellular distributions of PML RARα proteins were observed with immunofluorescence. Results 1.0 μmol/L of As 2O 3 did not induce cell apoptosis and differentiation, but it significantly inhibited the growth of K V sublines. As 2O 3 showed the similar but more potent effects in K PML and K PLZF sublines. 1.0 μmol/L As 2O 3 treatment for 3 days induced growth inhibition by 32%±3%, 57%±4% and 54%±6%, respectively in K V, K PML and K PLZF sublines. 1.0 μmol/L ATRA also exerted, to a less extent than As 2O 3, growth inhibitory effects in K V sublines, which became more obvious in K PML but not in K PLZF sublines. In addition, PML/PML RARα proteins were decreased and even disappeared in K V and K PML sublines with the treatment of 1.0 μmol/L As 2O 3 for 48 hours. Conclusion PML RARα and PLZF RARα markedly enhance growth inhibitory effects of As 2O 3 on K562 cells.
出处
《中华医学杂志》
CSCD
北大核心
2000年第4期297-300,共4页
National Medical Journal of China
基金
国家杰出青年科学基金! ( 3 972 5 0 11)
国家自然科学基金! ( 3 973 0 2 70 )
面上项目! ( 3 9670 3 2 9
3 9770 410 )
上海
