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为了更全面地了解nm23-H1在肺癌中发挥转移抑制的机理,用双向凝胶电泳技术比较人高转移大细胞肺癌细胞株(L9981)和转染nm23-H1基因的人大细胞肺癌细胞株(L9981-nm23-H1)间蛋白表达的差异.利用固相pH梯度双向凝胶电泳分离人高转移大细胞肺癌细胞株(L9981)和转染nm23-H1基因的人大细胞肺癌细胞株(L9981-nm23-H1)的总蛋白,用图像分析软件比较分析以识别细胞间的差异表达蛋白质.结果成功地获得了两株细胞蛋白组分辨率高、重复性好的双向凝胶电泳图谱.软件分析两种细胞的凝胶电泳图谱后发现,在相同分析条件下识别的蛋白质斑点数L9981为902±169个、L9981-nm23-H1为1160±212个.比较L9981和L9981-nm23-H1人大细胞肺癌细胞株的双向凝胶电泳蛋白质图谱后发现6个蛋白质点仅在L9981中有表达,17个蛋白质点仅在L9981-nm23-H1中有表达.此外,发现13个在两种细胞株中均存在,但表达量差异在2倍以上的蛋白质点(P<0.05).结果提示,nm23-H1基因转染引起人高转移大细胞肺癌细胞株蛋白质表达谱的变化,可能是其逆转肺癌侵袭转移的生物学基础.
To further understand the mechanism of nm23-H1 metastasis inhibition in lung cancer, two-dimensional gel electrophoresis was used to compare the expression of nm23-H1 gene between human highly metastatic large cell lung cancer cell line L9981 and nm23-H1 transfected human large cell lung cancer cell line L9981 -nm23-H1) in human lung cancer cell line L9981 and nm23-H1 transfected human lung cancer cell line L9981-nm23-H1 were detected by two-dimensional gel electrophoresis H1) were analyzed by image analysis software to identify the differentially expressed proteins between cells.Results Two-dimensional gel electrophoresis patterns with high resolution and good repeatability of the two cell protein groups were successfully obtained.Analysis of two kinds of cells The results of gel electrophoresis showed that the L9981 and L9981-nm23-H1 protein spots identified under the same conditions were 902 ± 169 and 1160 ± 212, respectively.Comparing the L9981 and L9981-nm23-H1 human large cell lung cancer cell lines Two-dimensional gel electrophoresis of protein profiles showed that six protein spots were only expressed in L9981, and 17 protein spots were only expressed in L9981-nm23-H1. In addition, 13 were found in both cell lines but expressed The difference is between 2 More protein spots (P <0.05). The results suggest that, nm23-H1 transfected gene cause human high-metastatic large cell lung cancer cell line protein expression profile change, may be the biological basis for the reversal of lung cancer invasion and metastasis.