苦参碱和川芎嗪抑制白血病细胞侵袭转移作用的实验研究

目的探讨苦参碱及川芎嗪对低氧(3%O2)培养条件下人B淋巴细胞白血病细胞株Raji细胞、人慢性髓系白血病细胞株K562细胞侵袭转移的抑制作用及机制。方法体外常规培养细胞,低氧环境下继续培养24 h后,分别加入终浓度为0、0.15、0.2、0.25 g/L苦参碱或0、0.1、0.15、0.2 g/L川芎嗪处理,以常氧培养不加药物的细胞为对照组,分别检测细胞黏附、迁移和侵袭能力,并以RT-PCR方法检测HIF-1α、VEGF mRNA的表达。结果低氧环境下Raji细胞和K562细胞的黏附、迁移和侵袭能力,以及HIF-1α、VEGF mRNA的表达均较常氧对照组显著增强(P<0.01)。0.15、0.2、0.25 g/L苦参碱均有效抑制低氧环境下Raji细胞的黏附、迁移和侵袭能力,并下调HIF-1α、VEGF mRNA的表达(P<0.05)。0.2、0.25 g/L苦参碱均有效抑制低氧环境下K562细胞的黏附、迁移和侵袭能力并下调HIF-1α、VEGF mRNA的表达(P<0.05)。0.1、0.15、0.2 g/L川芎嗪均有效抑制Raji细胞、K562细胞的黏附、迁移和侵袭能力,并下调HIF-1α、VEGF mRNA的表达(P<0.05),均呈剂量依赖性。结论苦参碱和川芎嗪能有效抑制低氧环境下Raji细胞和K562细胞黏附、迁移和侵袭能力,其作用机制可能通过下调细胞内HIF-1αmRNA的表达,从而减少VEGF mRNA的生成来实现。

Inhibitory effects of matrine and ligustrazine on invasion and metastasis of leukemia cells treated with hypoxia culture

Objective To study the inhibitory effects of matrine and ligustrazine on invasion and metastasis of leukemia cells treated with hypoxia.Methods Leukemia cell line Raji and K562 were cultured in vitro and treated with 3% oxygen for 24 hours.The cells were collected and treated with matrine(0,0.15,0.2,0.25g/L) or ligutrazine(0,0.1,0.15,0.2g/L).The cells cultured under normoxic and drug-free conditions were used as control.Cell adhesion,migration and invasion assays were performed to observe the effect of matrine or ligutrazine on the adhesion,migration and invasion in K562 and Raji cells.RT-PCR was used to evaluate the mRNA expression levels of HIF-1α and VEGF.Results Compared to the control group,the cells cultured under hypoxia and drug-free coditions could significantly enhance the cell adhesion,migration and invasion as well as the mRNA expression levels of HIF-1α and VEGF in both Raji and K562 cells(P<0.01);0.15,0.2 and 0.25 g/L matrine could significantly inhibit the cell adhesion,migration and invasion as well as the mRNA expression levels of HIF-1α and VEGF in Raji cells(P<0.05);0.2 and 0.25 g/L matrine could significantly inhibit the cell adhesion,migration and invasion as well as the mRNA expression levels of HIF-1α and VEGF in K562 cells(P<0.05);0.1,0.15 and 0.2 g/L lingutrazine could significantly inhibit the cell adhesion,migration and invasion as well as the mRNA expression levels of HIF-1α and VEGF in both Raji and K562 cells(P<0.05);All the changes were in a dose-dependent manner.Conclusions Both matrine and lingutrazine are good inhibitors of invasion and metastasis of leukemia cells under hypoxic conditions.Matrine and lingutrazin might decrease the mRNA expression level of VEGF via down-regulation of HIF-1α.