硼替佐米对伊马替尼耐药细胞株K562的凋亡作用及机制.PDFVIP

·1136 · doi 10.3969/j.issn. 1673-4254.2017.08.23 J South Med Univ, 2017, 37(8): 1136-1139 基础研究 硼替佐米对伊马替硼替佐米对伊马替尼耐药细胞株尼耐药细胞株KK562562 的凋亡作用及机制的凋亡作用及机制 1 2 2 2 华佳叶 ，周旭红 ，欧阳舒婷 ，吴泳彬 1 2 广州医科大学附属第二医院 血液科，血液实验室，广东 广州 510260 摘要：目的探讨硼替佐米对伊马替尼耐药细胞株K562（K562R）的诱导凋亡作用及机制。方法 采用浓度梯增法建立耐伊马替 尼K562 细胞系。应用CCK-8法检测不同浓度硼替佐米分别作用K562 和K562R 细胞24 h、48 h 后，对细胞的增殖抑制的效果。 使用流式细胞术方法分别检测硼替佐米单独或联合伊马替尼作用于K562R 细胞48 h 后的细胞凋亡。应用Western blotting 检 测不同浓度硼替佐米作用K562R 细胞48 h 后，Mcl- 1、Bcl-2、Bcr/Abl 蛋白表达的异同。结果 成功将对伊马替尼敏感的K562 细 胞诱导为K562R ，耐药倍数为31.8；硼替佐米对K562及K562R 细胞的增殖抑制呈浓度、时间依赖（P均0.05）；随着硼替佐米浓 度增加，对K562R 细胞的诱导凋亡作用增强，且硼替佐米与伊马替尼联合具有协同作用（P0.05）；硼替佐米可抑制K562R 细胞 Mcl- 1，Bcr/Abl 蛋白的表达，Bcl-2 无变化。结论 硼替佐米具有对K562R 细胞的增殖抑制和诱导凋亡作用，其机制可能与下调 K562R 细胞Bcr/Abl 和MCL- 1的表达以及促进MCL- 1发生切割有关。 关键词：硼替佐米；K562 细胞；抗药性；伊马替尼 Effect of bortezomib in inducing apoptosis of imatinib-resistant K562 cells and the mechanism 1 2 2 2 HUA Jiaye , ZHOU Xuhong , OUYANG Shuting , Wu Yongbin 1Department of Hematology, 2Hematology Laboratory, Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China Abstract: Objective To investigate the effect of bortezomib in inducing apoptosis in imatinib-resistant K562 (K562R) cells and its possible mechanism. Methods K562 cells were cultured in gradient concentrations of imatinib for several months to generate imatinib-resistant K562 cells. The viability of K562R cells treated with bortezomib was measured using CCK-8 cell proliferation assay, and the cell apoptosis was analyzed by flow cytometry with annexin V/PI dual staining. Western blotting was used to detect the protein expressions of Mcl-1,Bcl-2 and Bcr/Abl. Results K562R cell line was successfully established, which showed