小分子酪氨酸激酶抑制剂AG-490对人慢性髓系白血病急变细胞株K562细胞增殖凋亡的影响

Effects of Small Molecule Tyrosine Kinase Inhibitors AG-490 on Proliferation and Apoptosis of Chronic Myeloid Leukemia Cell Line K562 Cells

目的探讨酪氨酸激酶抑制剂AG-490对人慢性髓系白血病急变细胞株K562细胞的增殖及凋亡的影响。方法采用MTT法观察AG-490在体外对人慢性髓系白血病急变细胞株K562细胞增殖的影响,光学显微镜下观察AG-490对K562细胞形态的变化,流式细胞术检测AG-490对K562细胞凋亡及细胞周期的影响,逆转录-聚合酶链反应(RT-PCR)及实时荧光定量PCR技术检测AG-490作用后K562细胞bcr/abl融合基因及凋亡相关基因c-myc mRNA的表达水平。结果 AG-490可明显抑制K562细胞的增殖,并随药物作用时间的延长和浓度的增加,抑制作用越明显(P<0.05);AG-490可使细胞周期阻滞于G1期,促使其凋亡(P<0.05)。AG-490作用于K562细胞48 h后bcr/abl融合基因的表达无明显改变(P>0.05),而c-myc的表达水平明显降低(P<0.05)。结论 AG-490可能通过阻断JAK2信号通路而下调c-myc的表达,使K562细胞阻滞于G1期,从而抑制K562细胞增殖,促使其凋亡。

Objective To investigate the effects of tyrosine kinase inhibitor AG-490 on proliferation and apoptosis of chronic myeloid leukemia cell line K562 cells.Methods Chronic myelogenous leukemia cell line K562 cells were in vitro cultured and its proliferation inhibition were detected by MTT assay,K562 cells morphological changes were observed by microscopy,K562 cells apoptosis and cycle were tested by flow cytometry,the expression of bcr/abl mRNA was detected by reverse transcription-polymerase chain reaction（RT-PCR）,the expression of bcr/abl and c-myc mRNA were detected by real-time quantitative PCR（RQ-PCR）.Results AG-490 could significantly inhibit K562 cell proliferation,and this effect could be more obviously with the extension of time and the increase of concentration;AG-490 could arrest K562 cell cycle at G1 phase,and increase the apoptosis of cells（P0.05）.The expression of bcr/abl fusion gene had no changes（P0.05）,while the level of c-myc mRNA decreased significantly after treated by AG-490（P0.05）.Conclusion AG-490 could be down-regulate the level of expression of c-myc gene via blocking JAK2 signal transduction pathway to inhibit the proliferation of K562 cells and induce the apoptosis of K562 cells by arresting the cell cycle at G1 phase.