论文部分内容阅读
目的:膜研究膜型-1基质金属蛋白酶(MT1- MMP)反义RNA对人胃癌细胞BGC823靶基因表达和侵袭特性的影响方法:利用基因重组技术构建人MT1-MMP反义RNA真核表达载体,转染人胃癌细胞BGC823,应用RT-PCR、MTT、明胶酶谱和体外侵袭实验等方法观察人胃癌细胞BGC823转染前后,MT1-MMP mRNA表达水平、细胞生长、明教酶A活性及细胞体外侵袭能力等指标的变化.结果:成功构建了MT1-MMP反义RNA真核表达载体pasMMP14,将其转染胃癌细胞BGC823后,与阴性对照组相比,实验组MT1- MMP mRNA表达水平降低,抑制率为36%.转染48 h,明教酶A的活化受到了明显抑制.转染72 h,细胞增殖明显受抑(t=2.358,P<0.01 vs空白组:t=2.727 P<0.01 vs阴性组).实验组的穿膜细胞数明显低于空白对照组和阴性对照组(t=5.744,P<0.01;t=5.695,P<0.01).结论:反义RNA对人胃癌细胞MT1-MMP基因表达和侵袭能力具有明显的抑制作用,MT1-MMP基因可作为胃癌抗侵袭治疗的分子靶点.
AIM: To investigate the effect of membrane-1 matrix metalloproteinase (MT1-MMP) antisense RNA on the gene expression and invasion of human gastric cancer cell line BGC823.Methods: The eukaryotic expression vector of human MT1-MMP antisense RNA was constructed by gene recombination , Transfected human gastric cancer cell line BGC823, the expression of MT1-MMP mRNA, the growth of cells, the activity of inducible A (myo-oncoenzyme A) and the cell growth and proliferation of human gastric cancer cell BGC823 were observed by RT-PCR, MTT, gelatin zymography and in vitro invasion assay Invasiveness and so on.Results: The eukaryotic expression vector pasMMP14 of MT1-MMP antisense RNA was successfully constructed and transfected into BGC823 gastric cancer cells.Compared with the negative control group, the expression of MT1-MMP mRNA in the experimental group decreased, The inhibition rate was 36% .After 48 hours of transfection, the activation of the enzyme A was significantly inhibited. The proliferation of cells was significantly inhibited at 72 h after transfection (t = 2.358, P <0.01 vs control group: t = 2.727 P <0.01 vs (T = 5.744, P <0.01; t = 5.695, P <0.01) .Conclusion: Antisense RNA can inhibit the expression of MT1- MMP gene expression and invasion ability has a significant inhibitory effect, MT1-MMP gene can be used as gastric cancer resistance Molecular target therapy.