本文關(guān)鍵詞： 黃芩素 宮頸癌Hela 細(xì)胞 MMP2 MMP9 TIMP2 CyclinD1　出處：《瀘州醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：[目的]觀察不同濃度黃芩素干預(yù)宮頸癌Hela細(xì)胞后，細(xì)胞周期、細(xì)胞遷移、細(xì)胞培養(yǎng)基中基質(zhì)金屬蛋白酶2（matrix metalloproteinase2，MMP2）、基質(zhì)金屬蛋白酶9（matrix metalloproteinase，MMP9）活性的變化，以及MMP2、MMP9、基質(zhì)金屬蛋白酶組織抑制劑2（Tissue Inhibitor of Metalloproteinase2，TIMP2）、細(xì)胞周期蛋白D1（CyclinD1）mRNA和蛋白表達(dá)水平的變化。[方法]體外培養(yǎng)宮頸癌Hela細(xì)胞，分空白對照（Control）組、（50，100，200，300）μmol/L干預(yù)組，培養(yǎng)（12，24，48）h。相差顯微鏡下觀察細(xì)胞形態(tài)及數(shù)量的變化；流式細(xì)胞技術(shù)法檢測細(xì)胞周期的變化；細(xì)胞遷移實驗觀察細(xì)胞遷移的改變；明膠酶譜法檢測細(xì)胞培養(yǎng)基中的MMP2、MMP9活性變化；RT-PCR檢測MMP2、MMP9、TIMP2、CyclinD1mRNA表達(dá)水平的變化；Western Blot檢測MMP2、MMP9、TIMP2、CyclinD1蛋白表達(dá)水平的變化。[結(jié)果]黃芩素干預(yù)24小時后，與空白對照組相比較，各黃芩素干預(yù)組的細(xì)胞G1期均受阻滯，阻滯程度與黃芩素干預(yù)濃度成正向變化；黃芩素能有效抑制Hela細(xì)胞的遷移，遷移距離與黃芩素干預(yù)濃度成反向變化；細(xì)胞培養(yǎng)基中MMP2及MMP9的活性受抑制，，抑制程度與黃芩素干預(yù)濃度成正向變化；MMP2、MMP9、CyclinD1mRNA及蛋白表達(dá)隨黃芩素干預(yù)濃度增加，呈逐漸降低的趨勢（P0.05，P0.01）, TIMP2mRNA及蛋白水平表達(dá)隨黃芩素濃度增加而逐漸增強(qiáng)（P0.05，P0.01）。[結(jié)論]黃芩素可有效抑制宮頸癌Hela細(xì)胞的遷移，阻滯細(xì)胞周期中G1期、降低MMP2、MMP9活性、上調(diào)TIMP2及下調(diào)MMP2、MMP9、CyclinD1的表達(dá)，發(fā)揮抑制宮頸癌Hela細(xì)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.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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