苦参碱对HeLa细胞粘附和移动的抑制作用及机制

目的:探讨苦参碱(Matrine)抑制HeLa细胞粘附和移动的作用及机制。方法:不同浓度苦参碱作用HeLa细胞24h后,粘附实验检测细胞粘附率,划痕损伤实验检测迁移速度,Westernblotting检测血管扩张刺激磷蛋白(VASP)磷酸化水平,PKA试剂盒检测蛋白激酶A(PKA)活性。结果:5,50,100,250和500μg/ml苦参碱处理24h后,HeLa细胞粘附率分别为74.68%,48.97%,45.66%,40.68%和32.61%,总体呈下降趋势,50μg/ml组分别与5μg/ml和500μg/ml组比较差异有显著性(均为P<0.01);与100μg/ml和250μg/ml组比较差异无显著性(P>0.05)。50μg/ml苦参碱处理24h后HeLa细胞迁移速度明显降低(P<0.01),PKA活性明显降低(P<0.01),VASP处于非磷酸化状态。结论:苦参碱抑制HeLa细胞粘附和运动,并降低其PKA活性及抑制骨架调节蛋白VASP磷酸化,发挥了抑制肿瘤转移作用。

Inhibitive Effect and Mechanism of Matrine on Adhesion and Migration of HeLa Cells

Objective: To explore the inhibitive effect and mechanism of Matrine on adhesion and migration of HeLa cells. Methods: Adhesive ratio was detected by adhesion assay, migration rate was measured by wound healing assay, PKA activity was tested by PKA activity assay and phosphorylation level of VASP was detected by Western blotting. Results: After HeLa cells were treated with Matrine of 5, 50,100,250 and 500 μg/ml for 24 h, adhesive ratios were 74.68%, 48.97%,45.66%, 40.68% and 32.61%, respectively. Compared with that of the 5 μg/ml and 500 μg/ml groups, the adhesive ratio of 50 μg/ml Matrine group was significantly different(P<0.01), but compared with that of the 100 μg/ml group and 250 μg/ml group, the value of 50 μg/ml Matrine group had no significant difference(P>0.05). After HeLa cells were treated with 50 μg/ml Matrine for 24 h, the migration rate was markedly reduced (P<0.01), PKA activity also was significantly reduced (P< 0.01) and VASP showed no phosphorylation state.Conclusion: Matrine can inhibit adhesion and migration of HeLa cells as well as reduce their PKA activity and inhibit phosphorylation of VASP . Matrine may play anti-metastatic roles.