黄芩素抑制宫颈癌Hela细胞增殖作用机制的初步研究

发布时间：2018-01-27 08:09

本文关键词： 黄芩素宫颈癌Hela 细胞 MMP2 MMP9 TIMP2 CyclinD1　出处：《泸州医学院》2014年硕士论文　论文类型：学位论文

【摘要】：[目的]观察不同浓度黄芩素干预宫颈癌Hela细胞后，细胞周期、细胞迁移、细胞培养基中基质金属蛋白酶2（matrix metalloproteinase2，MMP2）、基质金属蛋白酶9（matrix metalloproteinase，MMP9）活性的变化，以及MMP2、MMP9、基质金属蛋白酶组织抑制剂2（Tissue Inhibitor of Metalloproteinase2，TIMP2）、细胞周期蛋白D1（CyclinD1）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、（50，100，200，300）μmol/L干预组，培养（12，24，48）h。相差显微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、TIMP2、CyclinD1mRNA表达水平的变化；Western Blot检测MMP2、MMP9、TIMP2、CyclinD1蛋白表达水平的变化。[结果]黄芩素干预24小时后，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、MMP9、CyclinD1mRNA及蛋白表达随黄芩素干预浓度增加，呈逐渐降低的趋势（P0.05，P0.01）, TIMP2mRNA及蛋白水平表达随黄芩素浓度增加而逐渐增强（P0.05，P0.01）。[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、上调TIMP2及下调MMP2、MMP9、CyclinD1的表达，发挥抑制宫颈癌Hela细胞增殖的作用。
[Abstract]:[Objective] to observe the cell cycle and cell migration after different concentrations of baicalin interfered with cervical cancer Hela cells. Matrix metalloproteinase 2 (MMP2) in cell culture medium. Changes of matrix metalloproteinase 9 (MMP9) activity and MMP2 / MMP9 activity. Tissue inhibitor of matrix metalloproteinase (tissue inhibitor of matrix metalloproteinase), tissue Inhibitor of metalloproteinase 2 (TIMP2). The changes of cyclin D _ 1 mRNA and protein expression in cyclin D _ 1. [Methods: cervical cancer Hela cells were cultured in vitro and divided into two groups: control group (control group) and control group (control group). The morphologic and quantitative changes of cells were observed under phase contrast microscope. The changes of cell cycle were detected by flow cytometry. Cell migration was observed by cell migration assay. The activity of MMP2 + MMP9 in cell culture medium was detected by gelatinase assay. RT-PCR was used to detect the expression of cyclin D1 mRNA in MMP2, MMP9 and TIMP 2. Western Blot was used to detect the expression of cyclin D1 in MMP2, MMP9 and TIMP 2. [Results after 24 hours of baicalin intervention, compared with the blank control group, the G1 phase of the cells in each baicalin intervention group was blocked, and the degree of block was positively changed with the concentration of baicalin intervention. Baicalin could effectively inhibit the migration of Hela cells, and the migration distance changed inversely with the intervention concentration of baicalin. The activities of MMP2 and MMP9 in cell culture medium were inhibited and the inhibition degree was positively correlated with the intervention concentration of baicalin. The expression of CyclinD1 mRNA and protein in MMP2 + MMP9 was decreased gradually with the increase of baicalin concentration. The expression of TIMP2mRNA and protein increased with the increase of baicalin concentration. [Conclusion: baicalin can effectively inhibit migration of cervical cancer Hela cells, block G1 phase of cell cycle, decrease MMP2mMP9 activity, up-regulate TIMP2 and down-regulate MMP2mMP9. The expression of CyclinD1 inhibits the proliferation of cervical cancer Hela cells.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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