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观察内源性超氧阴离子自由基对癌基因表达的影响．方法：构建正、反义人MnSOD（SOD2）真核表达载体，转入食管瘤细胞Eca-109；通过升高或降低细胞内SOD2的水平来增加或减少细胞内O2-的清除从而改变细胞内O2-的水平；以RNA斑点杂交和免疫细胞化学方法检测癌基因表达的变化，以流式细胞仪检测细胞周期变化．结果：基因转入细胞并表达．转染SOD2基因细胞内SOD2水平升高，Cu，Zn-SOD（SOD1）水平未变，细胞内O2-水平下降49％以上；bcl-2表达下调，p53，c-Ha-ras表达轻度上调；流式细胞仪检测S期细胞减少．转染反义SOD2细胞SOD2水平降低，但SOD1活力水平却升高，因而总SOD水平增加，细胞内O2-水平也降低32％以上；bcl-2，p53，ras表达均上调，S期细胞数变化不明显．结论：①通过转SOD2基因改变细胞内O2-水平的方法是可行的，但仍需改进；②细胞内O2-水平的变化可影响癌基因的表达，而且由转染SOD2基因引起细胞内O2-水平降低对Eca-109食管癌细胞的增殖表现出抑制作用．
Observe the effect of endogenous superoxide anion radical on the expression of oncogene. METHODS: Positive and negative human MnSOD (SOD2) eukaryotic expression vectors were constructed and transfected into esophageal carcinoma cells Eca-109. O2 clearance was increased or decreased by increasing or decreasing intracellular SOD2 levels. The level of O2- was detected by RNA dot blot hybridization and immunocytochemistry, and cell cycle changes were detected by flow cytometry. Results: Genes were transferred to cells and expressed. The levels of SOD2 in SOD2 transfected cells were increased, the levels of Cu, Zn-SOD (SOD1) were unchanged, and the intracellular O2-level was decreased by 49%; the expression of bcl-2 was down-regulated and the expression of p53, c-Ha-ras was slightly increased. Flow cytometry detection of S phase cytopenia. The level of SOD2 in transfected antisense SOD2 cells was decreased, but the SOD1 activity was increased, so the total SOD level was increased, the intracellular O2-level was also reduced by more than 32%; the expression of bcl-2, p53, ras was up-regulated, and the number of S phase cells was increased. The change is not obvious. Conclusions: 1 The method of changing the intracellular O2-level by SOD2 gene transfection is feasible, but still needs improvement; 2 The change of intracellular O2- level can affect the expression of oncogene, and the intracellular O2- induced by transfection of SOD2 gene The decrease in the level of Eca-109 esophageal cancer cells showed inhibition of proliferation.