胡桃醌对人宫颈癌HeLa细胞侵袭与迁移的影响

Effect and its mechanism of Juglone on invasion and metastasis ability of human cervical cell line HeLa

目的观察胡桃醌对HeLa细胞侵袭及迁移的抑制作用,并探讨其可能的作用机制。方法常规体外培养人HeLa细胞,加入10、20、50、100μmol/L浓度的胡桃醌,继续培养24 h。通过倒置显微镜观察细胞形态学变化,采用MTT法检测细胞增殖情况,细胞划痕实验检测胡桃醌对细胞迁移能力的影响,用Transwell小室法检测细胞侵袭能力的影响。Western blotting检测基质金属蛋白酶(MMP)-2、MMP-9蛋白的表达。结果细胞形态学观察显示,不同浓度胡桃醌均可导致细胞形态学发生改变;MTT检测表明,胡桃醌对HeLa细胞的生长有明显抑制作用,且抑制作用随药物浓度的增加而增强(P<0.05或P<0.01);划痕实验结果显示,随胡桃醌药物浓度的增加,细胞的平面运动能力明显下降;Transwell侵袭小室实验结果显示,胡桃醌对HeLa细胞体外的侵袭力具有很强的抑制作用;Western blotting结果表明,胡桃醌给药能够抑制HeLa细胞MMP-2、MMP-9蛋白的表达。结论胡桃醌能够抑制HeLa细胞的增殖并抑制细胞侵袭,其机制可能与抑制MMP-2、MMP-9的表达有关。

Objective To observe the effect of Juglone on invasion and metastasis of Hela cells and explore the possible mechanism. Methods HeLa cells were cultured and treated with 10,20,50,100μmol/L Juglone for 24 hours.The morphology changes of HeLa cells were observed with an inverted microscope. The viability of HeLa cells was detected by MTT assay. The cell scratch test was used to detect cell migration ability after treatment of Juglone. The ability of cell invasion was measured by Transwell chamber. The expression of matrix metalloprateinases（ MMP）-2 and MMP-9 were detected by Western blotting. Results Compared with control group,the viability of HeLa cells decreased after treatment with different concentrations of Juglone for 24 hours,and the cell morphology was changed in a dose-dependent manner.Scratch test results showed that the level of cell movement ability decreased significantly with the increase of the concentration of Juglone. Transwell invasion assay showed that Juglone had a strong inhibitory effect on the invasiveness of HeLa cells in vitro. Western blotting results showed that Juglone inhibited the expression of MMP-2 and MMP-9 protein in HeLa cells. Conclusion Juglone can inhibit the invasion and metastasis in HeLa cells,and its possible mechanism may be related to down regulating the expression of MMP-2 and MMP-9.