CD44v6对急性白血病细胞HL-60和THP-1增殖和迁移的影响

目的:探讨CD44变异亚型对急性白血病细胞增殖和迁移的影响。方法:选择对数生长期的急性白血病细胞株HL-60、THP-1和慢性白血病细胞株K562,采用荧光定量PCR法检测CD44v6mRNA的表达。通过电转的方法转染CD44v6siRNA到HL-60和THP-1细胞抑制细胞的CD44v6表达,通过western方法检测CD44v6蛋白的抑制情况。将实验分成HL-60+N-siRNA、HL-60+CD44V6-siRNA、THP-1+N-si RNA、THP-1+CD44V6-siRNA共4组,培养24、48、72 h后分别取细胞悬液用台盼蓝染色后计数活细胞数检测细胞的增殖情况;使用Transwell小室培养法观察HL-60和THP-1细胞的迁移率。结果:通过荧光定量PCR方法检测THP-1和HL-60细胞均高表达CD44v6(分别为0.0037±0.0007和0.00292±0.0002),明显高于K562的表达(P0.01);转染后的HL-60和THP-1细胞株中CD44v6蛋白表达水平明显下调,细胞计数结果显示转染CD44v6-siRNA的HL-60和THP-1细胞在24、48和72 h增殖均明显下降。迁移实验结果显示THP-1+N-si RNA和HL-60+N-si RNA细胞的迁移率为17%和23%,与相应对照组相比THP-1+CD44v6-siRNA和HL60+CD44v6-siRNA组细胞24 h迁移率明显下降(分别降至11%和14%)。结论:CD44v6可以通过干预白血病细胞的增殖和迁移能力,参与调解白血病细胞的增殖和髓外进展。

The Influence of CD44v6 on Proliferation and Migration in HL-60 and THP-1 Cells

ABSTRACT Objective: To investigate the influence of CD44v6 on proliferation and migration in HL-60 and THP-1 cells. Methods: The expression levels of CD44 in leukemia cell lines, when they were in logarithmic phaseTHP-1, HL-60 and K562 were assayed by qRT-PCR. CD44v6 siRNA was transfected into THP-1and HL-60 cell lines by electransfection. And these cell lines were divided into control group (transfected with N-siRNA) and experimental group (transfected with siRNA-CD44V6). Western blot was used to measure the protein expression of CD44v6 in the experimental and the control groups. The proliferation and migration of cells were measured by cell count and the artificial matrix membrane. Results: The relative expression ratios of CD44 to GAPDH in THP-1 and HL-60 were 0.0037±0.0007 and 0.00342±0.0005, respectively, which were significantly higher than that in K562 cell (P<0.01). After THP-1 and HL-60 cells were transfected with siRNA-CD44v6 inhibit the protein expression of CD44v6. Compared with the control group, the experimental group had a significant decrease in proliferative capacity at 24, 48, 72 h. Migration assay showed that the transmembrane rates of THP-1+N-siRNA and HL-60+N-siRNA cells were 17% and 23% respectively, and decreased to 11% and 14% in HL60+ CD44V6-siRNA and THP-1+ CD44V6-siRNA groups respectively. Conclusion: CD44V6 might play an important role in the migration and proliferation of leukemia cells and be involved in the extramedullary infiltration of leukemia cells.