苦参碱对宫颈癌HeLa细胞黏附和侵袭的抑制作用及其机制

Effect and mechanism of matrine on adhesion and invasion of cervical cancer He La cells

目的观察苦参碱对宫颈癌He La细胞黏附和侵袭转移的抑制作用及其可能机制。方法不同浓度苦参碱(0,0.2,0.4,0.6,0.8,1.0和1.5 g/L)干预He La细胞后,采用MTT法检测苦参碱对He La细胞增殖的影响。不同浓度苦参碱(0,0.2,0.4和0.6 g/L)干预He La细胞后,细胞黏附试验和Transwell小室法分别检测苦参碱对He La细胞黏附能力和侵袭能力的影响。利用实时定量PCR和免疫印迹检测苦参碱对MMP-2和MMP-9 mRNA和蛋白表达水平的影响。免疫印迹法检测苦参碱对p38和p-p38蛋白表达水平的影响。结果当浓度超过0.8 g/L时,苦参碱能够明显抑制He La细胞的增殖(P<0.05)。在低于增殖抑制浓度0.8 g/L时,苦参碱也能够有效抑制He La细胞的黏附和侵袭(P<0.05),这种作用具有明显的剂量依赖性。当浓度超过0.2 g/L时,苦参碱能够有效抑制MMP-2的转录和MMP-9蛋白的表达;当浓度高于0.4 g/L时,苦参碱能够有效抑制He La细胞中MMP-9的转录和MMP-2蛋白的表达(P<0.05),具有明显的剂量依赖性。进一步的检测发现苦参碱在浓度超过0.4 g/L时,能够有效抑制He La细胞中p38的磷酸化水平(P<0.05)。结论苦参碱能够有效抑制He La细胞的黏附和侵袭,这种作用可能与其通过抑制p38信号通路的活性进一步调节MMP-2和MMP-9的表达有关。

Objective To explore the anti-metastatic effect of matrine and its mechanism in human cervical cancer He La cells.Methods He La cells were treated with different concentrations of matrine（ 0,0. 2,0. 4,0. 6,0. 8,1. 0 and 1. 5 g / L）,then the inhibitory effect of matrine on the growth of He La cells was measured by MTT assay. After the He La cells were treated with different concentrations of matrine（ 0,0. 2,0. 4 and 0. 6 g / L）,the cell adhesion and the invasion were analyzed by MTT and Transwell assay. Moreover,real time PCR and Western blot were used for investigating the expression of MMP-2,MMP-9,p38 and p-p38. Results Matrine significantly inhibited the proliferation of He La cells in the concentration of above 0. 8 g / L（ P 〈 0. 05）. Matrine inhibited the adhesion and invasion ability of He La cells in a concentration-dependent manner in the concentrations of below 0. 8 g / L（ P 〈 0. 05）. Matrine effectively down-regulated the transcription of MMP-2 mRNA and the expression of MMP-9 protein in the concentration of above 0.2 g / L,and down-regulated the transcription of MMP-9 mRNA and the expression of MMP-2 protein in the concentration of above 0. 4g / L in a concentration-dependent manner（ P 〈 0. 05）. Matrine also significantly down-regulated the phosphorylation level of p38 in the concentration of above 0. 4 g / L（ P 〈 0. 05）. Conclusion These results suggest that matrine could induce the adhesion and the invasion of He La cells,which may be related to the inhibition of MMP-2 and MMP-9 in He La cells through the p38 signal pathway.