尼洛替尼诱导K562/A02细胞凋亡及对HO-1基因表达的影响

 目的 研究尼洛替尼(nilotinib，AMN107诱导K562/A02细胞凋亡及对血红素加氧酶-1(HO-1基因表达的影响。方法 采用荧光原位杂交(FISH法检测K562/A02细胞中的BCR-ABL融合基因；用不同浓度的AMN107分别处理K562/A02细胞24 h后，通过实时荧光定量PCR(RQ-PCR法检测BCR-ABL融合基因mRNA的表达水平；采用MTT法观察细胞增殖变化；通过Annexin V/PI双染色法检测细胞凋亡和细胞周期；采用RT-PCR法和Western Blot法检测HO-1基因的表达。 结果 FISH法分析结果显示，K562/A02细胞BCR-ABL融合基因阳性细胞占细胞总数98%；RQ-PCR法检测结果显示，0，5，10，20 μmol·L-1 AMN107作用于K562/A02细胞24 h后BCR-ABL融合基因表达随AMN107浓度增加而逐渐下降；MTT法和Annexin V/PI法显示与对照组相比，细胞存活率随AMN107浓度升高而下降，凋亡率逐渐增加；细胞周期分析显示，经AMN107处理的细胞，G₀/G₁期和S期细胞明显减少，细胞阻滞在G₂/M期；RT-PCR法和Western blot法均检测到HO-1基因表达随AMN107浓度升高而降低。结论 AMN107可抑制BCR-ABL融合基因和HO-1基因表达，从而诱导慢性粒细胞白血病（CML耐药细胞的凋亡，说明HO-1是CML细胞生长以及BCR-ABL基因存在的一个相关因子，HO-1基因是克服慢性粒细胞白血病耐药的一个潜在靶向。

Effect of Nilotinib on HO-1 Gene Expression and Induction of Apoptosis in K562/A02 Cells

OBJECTIVE To investigate the effect of nilotinib on HO-1 gene expression and induction of apoptosis in K562/A02 cells.METHODS BCR-ABL in K562/A02 cell was detected by Fluorescence in situ hybridization(FISH).K562/A02 cells were treated with AMN107 of different concentrations for 24 h.The expression of BCR-ABL gene on mRNA level was detected by RQ-PCR.Cell proliferation was observed by MTT.Cell apoptosis and cell cycle were inspected by Annexin V/PI double staining method.The expression of HO-1 gene was dete...