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目的 探讨bcl 2基因在急性白血病细胞凋亡及耐药性中的作用及Bcl 2反义寡核苷酸(ASON)逆转耐药性的效果。方法 细胞耐药性测定采用MTT法 ,Bcl 2和Bax蛋白含量测定采用单克隆抗体标记技术在流式细胞仪上测定 ,细胞凋亡率采用流式细胞仪测定。结果 (1)Bcl 2水平在HL 6 0 HHT细胞系明显高于HL 6 0细胞系 (P <0 0 1) ;(2 )Bcl 2ASON可逆转HL 6 0 HHT的耐药性 (P<0 0 1) ;(3)HL 6 0 HHT细胞系的凋亡百分率经ASON作用明显升高 (P <0 0 5 ) ,经ASON +HHT作用后升高更加显著 (P <0 0 1) ;(4)Bcl 2ASON可明显降低Bcl 2表达水平 (P <0 0 1)。结论 在HL 6 0 HHT细胞系中 ,Bcl 2的高度表达是造成其耐药的重要原因 ;反义技术可以通过下调Bcl 2 Bax比值 ,部分逆转它的耐药的发生。通过进一步的深入研究 ,反义技术将有可能应用于难治及耐药的急性白血病。
Objective To investigate the role of bcl 2 gene in apoptosis and drug resistance of acute leukemia cells and the effect of Bcl 2 antisense oligonucleotide (ASON) in reversing drug resistance. Methods MTT assay was used to determine the drug resistance of the cells. The content of Bcl 2 and Bax protein was measured by flow cytometry using monoclonal antibody labeling technique. The apoptosis rate was determined by flow cytometry. Results (1) Bcl 2 level was significantly higher in HL 60 HHT cell line than in HL 60 cell line (P <0 01); (2) Bcl 2 ASON could reverse HL 60 HHT resistance (P < 0 0 1) (3) The percentage of apoptotic cells in HL 60 HHT cell line was significantly increased by ASON (P <0 05), and increased significantly after ASON + HHT (P <0 01); (4) Bcl 2ASON significantly reduced Bcl 2 expression (P <0 01). Conclusion The high expression of Bcl 2 in HL 60 HHT cell line is an important cause of its resistance. Antisense technology can partially reverse its resistance by down-regulating Bcl 2 Bax ratio. Through further in-depth research, antisense technology will likely be applied to refractory and resistant acute leukemia.