HFCL细胞诱导HL-60细胞分化和改变其基因表达谱的研究

本研究探索正常人骨髓成纤维细胞样基质细胞（HFCL）对急性髓系白血病HL-60细胞增殖分化影响的分子机理。采用流式细胞仪检测细胞周期,NBT还原实验和CD11b、CD14、CD13、CD33细胞表面抗原的测定检测细胞的分化;应用基因芯片技术检测HL-60细胞与HFCL细胞共培养前后基因表达谱的改变,并采用半定量RT-PCR、Northernblot对芯片结果进一步验证。结果发现,HL-60细胞与HFCL细胞共培养后,G0/G1期细胞明显增多,S期细胞减少;NBT阳性细胞增高,CD11b和CD14的表达增多,并有明显的统计学意义;基因表达谱改变的研究发现,在与HFCL细胞共培养后,HL-60细胞中582个基因表达上调,1323个基因表达下调。结论：HFCL细胞能诱导HL-60细胞部分分化,并能明显改变HL-60细胞的基因表达谱,为研究基质细胞与白血病细胞之间的相互关系及重要信号分子传导途径或cross-talk等提供了新的线索。

Differentiation of HL-60 Cells Directly Cocultured with HFCL Cells and Alteration of Their Gene Expression Profile

To study the molecular mechanism of the effect of fibroblastoid stromal cells(HFCL)from human bone marrow on the proliferation and differentiation in acute myeloid leukemia HL-60 cells,the cell cycle was analyzed by flow cytometry(FCM);the cell differentiation was determined by morphology NBT test and flow cytometric detection for expression of CD11b,CD14,CD13 and CD33;the genes differently expressed between HL-60 cells and HL-60 cells directly cocultured with HFCL were detected by using Affymetric oligo microarray technique.The changes of expression in some key genes were confirmed by using RT-PCR and Northern blot.The results showed that the percentage of G1 phase cells in AML cells cocultured with HFCL cells was higher than that without HFCL cells,and the percentage of S phase cells was lower.The NBT positive cells and the expression of CD11b and CD14 increased.It was found that after direct contact of HL-60 cells with HFCL cells for 96 hours,the expression levels of 582 genes were up-regulated,1 323 genes down-regulated.It is concluded that many genes may take part in the influence of HFCL cells on HL-60 cells,which may give important insights into the important molecules and pathways or cross-talk involved in the interaction between the AML cells and stromal cells.