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目的:研究重组反义c-myc腺病毒(Ad-AS-c-myc)诱导人急性早幼粒HL-60细胞系粒系分化及作用分子机制。方法:采用重组腺病毒Lac-Z(Ad-LacZ)及Ad-AS-c-myc联合硫酸鱼精蛋白转染HL-60细胞,X-gal染色判断转染效率。采用形态学、RT-PCR、流式细胞仪、四唑氮蓝(NBT)还原试验、非特异酯酶染色等方法进行研究。结果:Ad-AS-c-myc能抑制HL-60细胞c-myc基因在转录水平的表达。被转染HL-60细胞在形态上趋向成熟粒细胞,Ad-AS-c-myc使HL-60细胞周期G0/G1阻滞,过氧化物酶、非特异性酯酶活性增强。结论:Ad-AS-c-myc对HL-60细胞具有诱导粒系分化作用。其生物学作用与HL-60细胞c-myc的表达受抑、HL-60细胞出现一些成熟粒细胞的生理功能有关。
AIM: To investigate the molecular mechanism of granulocytic differentiation induced by recombinant antisense c-myc adenovirus (Ad-AS-c-myc) in human acute promyelocytic HL-60 cell line. Methods: HL-60 cells were transfected with recombinant adenovirus Lac-Z (Ad-LacZ) and Ad-AS-c-myc combined with protamine sulfate. X-gal staining was used to determine the transfection efficiency. The morphological, RT-PCR, flow cytometry, tetrazolium nitrogen blue (NBT) reduction test, non-specific esterase staining and other methods were studied. Results: Ad-AS-c-myc inhibited the expression of c-myc gene at the transcriptional level in HL-60 cells. Transfection of HL-60 cells in the morphology of mature granulocytes, Ad-AS-c-myc HL-60 cell cycle G0 / G1 block, peroxidase, non-specific esterase activity increased. Conclusion: Ad-AS-c-myc induces granulocytic differentiation in HL-60 cells. The biological role of HL-60 cells c-myc expression suppression, HL-60 cells appear some of the physiological function of mature granulocytes.