【作者】 曾维忠；

【导师】 周锐；

【作者基本信息】 中南大学 ， 呼吸内科， 2008， 硕士

【摘要】 第一部分靶向MMP-9的脱氧核酶抑制人肺腺癌A549细胞中MMP-9蛋白表达目的探讨靶向MMP-9的脱氧核酶(DNAzyme,Deoxyri bozyme)对人肺腺癌A549细胞中MMP-9蛋白表达的影响。确定靶向MMP-9的脱氧核酶对MMP-9的mRNA切割的有效性。方法设计靶向MMP-9的脱氧核酶,应用oligofectamine将脱氧核酶转染到A549细胞中。实验分三组:空白组为A549培养细胞,对照组为将control oligo转染A549细胞,实验组为靶向MMP-9的脱氧核酶转染A549细胞。靶向MMP-9的脱氧核酶与对照control oligo终浓度均为2 uM。采用Western Blot方法检测细胞中MMP-9蛋白表达。结果在靶向MMP-9的脱氧核酶干预1d后人肺腺癌A549细胞中MMP-9蛋白表达较对照组与空白组显著减低(p＜0.01),而空白组与对照组细胞中蛋白条带灰度值无明显差异(p＞0.05)。结论靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效地抑制MMP-9蛋白的高表达。第二部分靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响目的在证实靶向MMP-9的脱氧核酶能够有效抑制A549细胞中MMP-9蛋白表达的基础上,我们进一步研究靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响。方法应用oligofectamine将脱氧核酶转染到A549细胞中。利用细胞粘附与迁移试验观察脱氧核酶干预后细胞粘附与迁移能力的变化。结果经靶向MMP-9的脱氧核酶干预后,2h和4h细胞粘附率,迁移速度均显著降低(p＜0.01),而对照组与空白组之间无差异(P＞0.05)。结论靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效阻止细胞的粘附与迁移。更多还原

【Abstract】 ObjectiveTo investigate the effect of targeting MMP-9 deoxyribozyme on expression of MMP-9 in human lung adenocarcinoma cell line A549, then determine the effectiveness of targeting MMP-9 deoxyribozyme on cleavage of mRNA in MMP-9.MethodsDesigned the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. Experiment were divided into 3 groups: empty group: The human lung adenocarcinoma cell line A549. Control group: Transfected control oligo into human lung adenocarcinoma cell line A549. Experimental group: Tansfected targeting MMP-9 deoxyribozyme into human lung adenocarcinoma cell line A549. The concentrations of targeting MMP-9 deoxyribozyme and control oligo are 2 umol/L. Using western blot method to detect the expression of MMP-9 in cell.ResultsAfter targeting MMP-9 deoxyribozyme intervention one day, the expression of MMP-9 in the cells compared with the control group was significantly lower (p<0.01). There was no difference between the empty group and the control group in gradation of protein strap (p>0.05).ConclusionsTargeting MMP-9 deoxyribozyme effectively inhibited the expression of MMP-9 in human lung adenocarcinoma cell line A549. ObjectiveTo investigate the effect of targeting MMP-9 deoxyribozyme on cell adhesion and migration in human lung adenocarcinoma cancer cell line A549.MethodsDesigned the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. To observa the cell adhesion and migration after the intervention of deoxyribozyme.ResultsAfter targeting MMP-9 deoxyribozyme interventioned, the rate of cell adhesion and migration were significantly decreased (p<0.01). There was no difference between the empty group and the control group (p<0.05).ConclusionsTargeting MMP-9 deoxyribozyme effectively prevent cell adhesion and migration.更多还原