DADS抑制JAK1/STAT3信号通路诱导人白血病HL-60细胞分化

Induction of differentiation by diallyl disulfide through inhibition of JAK1/STAT3 in human leukemia HL-60 cells

目的探讨JAKs/STATs信号转导通路在二烯丙基二硫(DADS)诱导人白血病HL60细胞分化中的变化及其调控机制。方法将HL60细胞与DADS或JAKs/STATs信号通路的激酶抑制剂AG490在体外共同培养,观察细胞形态变化,检测药物作用前后细胞NBT还原能力及细胞表面分化抗原CD11b的改变;用Westernblot检测JAKs,STATs各家族成员在DADS诱导HL60细胞分化中的改变;并用免疫细胞化学法检测核转录基因STATs,cmyc,cfos,cjun的表达变化。结果DADS和AG490均可诱导HL60细胞向成熟粒系分化,且DADS在1.25mg·L-1时诱导分化作用达峰值;Westernblot检测JAK1,STAT3的酪氨酸激酶发生了磷酸化改变;免疫细胞化学示STAT3与cmyc基因蛋白核内表达下降,cjun,cfos基因蛋白核内表达上升。结论JAK1,STAT3酪氨酸激酶的磷酸化抑制参与了DADS诱导HL60细胞分化的调控,其机制可能通过调控与HL60细胞增殖分化相关的基因表达,抑制细胞DNA合成,从而抑制细胞增殖,诱导分化。DADS的作用相当于JAK1/STAT3信号通路的阻断剂。

Aim To investigate JAKs/STATs signal transduction change in HL-60 cells differentiation induced by diallyl disulfide(DADS)and molecular mechanism regulating the differentiation.Methods After incubation of HL-60 cells with DADS or AG490(50 μmol·L -1),the cell differentiation indexes were observed by cytomorphology, NBT reduction ability assay,cell myeloid differentiation antigen CD11b by flow cytometry. Kinase activity of JAKs/STATs was tested by western-blotting and expressions of nucleus transcription genes stats,c-myc,c-fos,c-jun were detected by immumocyte chemistry method.Results Cell differentiation index changes indicated that HL-60 cells were induced differentiation toward granulocytic lineage by DADS, Western blot test demonstrated that constitive phosphorylation of Jak1,stat3 kinase was suppressed. Stat3,c-myc gene expression decreased and c-fos, c-jun gene expression increased in HL-60 cells treated with DADS through immunocyte chemistry.Conclusion Inhibition of phosphative Jak1, Stat3 was involved in HL-60 cells differentiation induced by DADS, its molecular mechanism might be related to modulation of gene expression associated proliferation and differentiation,and inhibition of DNA systhesis, induction differentiation.