黄芩素抑制宫颈癌HeLa细胞迁移作用机制的研究

Experimental study of mechanism of baicalein inhibiting cervical cancer HeLa cell migration

目的:观察不同浓度黄芩素干预宫颈癌HeLa细胞后,细胞迁移、细胞培养基中基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)、基质金属蛋白酶9(matrix metalloproteinase,MMP9)活性的变化,以及MMP2、MMP9 mRNA和蛋白表达水平的变化。方法:体外培养宫颈癌HeLa细胞,分空白对照(Control)组、50、100、200、300μmol/L干预组,培养12、24、48 h。镜下观察细胞形态及数量的变化;流式细胞技术法检测细胞周期的变化;细胞迁移实验观察细胞迁移的改变;明胶酶谱法检测细胞培养基中的MMP2、MMP9活性变化;RT-PCR检测MMP2、MMP9 mRNA表达水平的变化;Western Bloting检测MMP2、MMP9蛋白表达水平的变化。结果:黄芩素干预24 h后,与空白对照组相比较,各黄芩素干预组的细胞G1期均受阻滞,阻滞程度与黄芩素干预浓度成正向变化;黄芩素能有效抑制HeLa细胞的迁移,迁移距离与黄芩素干预浓度成反向变化;细胞培养基中MMP2及MMP9的活性受抑制,抑制程度与黄芩素干预浓度成正向变化;MMP2及MMP9 mRNA及蛋白表达随黄芩素干预浓度增加,呈逐渐降低的趋势(P<0.05,P<0.01)。结论:黄芩素可有效抑制宫颈癌HeLa细胞的迁移,阻滞细胞周期中G1期、降低MMP2、MMP9活性,下调MMP2和MMP9的表达,发挥抑制宫颈癌HeLa细胞增殖的作用。

Objective： To observe the changes of migration and expression of mRNA and protein of cell matrix metal loprote-inase2（MMP2） and matrix metalloproteinase 9（MMP9） of HeLa cells under the intervention of baicalein.Methods： HeLa cells were cultured in vitro and were exposed to 0,50,100,200 and 300 μmol/L baicalein for 12,24,48 h.The motphological changes and cell number were detected by flow cytometry,the activity of MMP2/9 was detected by Gelatin Zymography,RT-PCR was used to measure the mRNA changes of MMP2/9 and Western Bloting was used to measure the protein expression of MMP2/9. Results： Compared to control group, baicalein significantly blocked cell cycle at G1,decreased the activity of MMP2/9 and decreased the mRNA and protein expression of MMP2/9 in a concentration-dependent manner（P 〈0.05,P 〈0.01）.Conclusion：Baicalein induces inhibition of HeLa cell growth by bloking cell cycle at G1 and decreasing the activity of MMP2/9,as well as down-regulate the mRNA and protein expression of MMP2/9.