糖基化磷脂酰肌醇特异性磷脂酶D对白血病细胞黏附和侵袭功能的影响

The role of Glycosylphosphatidylinositol-phospholipase D on the adhesiveness and invasiveness in leukemic cells

目的:探讨糖基化磷脂酰肌醇特异性磷脂酶D(GPI-PLD)对髓性白血病细胞黏附和侵袭功能及GPI锚定蛋白uPAR表达的影响。方法:二氮杂菲下调细胞的GPI-PLD酶活性;黏附和体外穿膜实验检测细胞的黏附、侵袭性;流式细胞术检测细胞膜uPAR表达;RT-PCR和Westernblot法检测THP-1细胞中GPI-PLD的表达水平。结果:二氮杂菲处理4h后,THP-1细胞的黏附率和膜uPAR的表达明显上调,差异有显著性;而细胞GPI-PLD的表达并无明显改变。结论:GPI-PLD可能参与THP-1细胞的黏附和侵袭,其机制可能与对uPAR表达的调节有关。

Objective To study the effects of GPI-PLD on the cleavage of membrane-bound uPAR and elucidate the implication for adhesive and invasive potential in myeloid leukemic cells. Method The various myeloid leukemic cells were treated with 1,10-phenanthroline, an inhibitor of GPI-PLD. Adhesion and invasion assays were applied to observe the adhesiveness and invasiveness of these cells, respectively. RT-PCR was used to detect the mRNA expression of GPI-PLD and uPAR in these cells. Also, we measured the expressive levels of active GPI-PLD protein by western blot assay. Simultaneously, the expression of membrane GPI-anehored uPAR on these cells was analyzed by flow cytometry. Results After incubated for 4 hours with 1,10-phenanthroline,the upregulations of adhesiveness and membrane uPAR expression were only observed in THP-1 ceils. However, no obvious changes of GPI-PLD expression were observed in these cells. Conclusion GPI-PLD may play an important role in the adhesion and invasion of THP-1 cells by regulating the expression of membrane GPI-anehored uPAR.