【摘要】 为探讨ω- 6脂肪酸脱氢酶基因fat -1在人类乳腺癌细胞MCF- 7中表达和对其生长的作用,将fat -1基因插入到腺病毒载体中,构建腺病毒重组载体(Ad·GFP·fat1) .通过包装细胞系(2 93)产生重组腺病毒,感染MCF 7细胞.用核糖核酸酶保护性分析技术,检测fat -1基因在MCF- 7细胞内的表达,细胞增殖试剂盒(MTT)和凋亡染色试剂盒染色分析fat 1基因对MCF- 7细胞增殖和凋亡的影响,用酶联免疫分析花生酸类(eicosanoids)前列腺素E2 (prostaglandinE2 )的含量.结果显示,腺病毒介导的fat- 1基因能在MCF- 7细胞内有效异源表达,抑制MCF -7细胞的增殖且导致凋亡,前列腺素的含量也明显地减少.结果说明,fat- 1基因在乳腺癌的基因治疗中具有良好利用价值.更多还原

【Abstract】 To explore transformation of fat-1 gene encoding an ω-6 fatty acid desaturase into human breast cancer cell to inhibit the proliferation of the cells,the fat-1 cDNA was cloned into pAd-CMV vector to construct a recombinant adenovirus and transfected 293 cells to obtain Ad·GFP·fat-1 recombinant virus.Immunofluorescence microscopy found that positive green signal was appeared in 60%～70% of total cells after two days of infection of human breast cancer cell line MCF-7 with the recombinant virus,which was further confirmed at mRNA level by ribonuclease protection assay.At the same time,the proliferation of MCF-7 cells was significantly attenuated to 80% of control group as assessed by MTT assay.The underlying mechanism for reduction of cell growth may due to increase of apoptotic rate induced by the fat-1 gene expression:Hoechst dye staining following by showed that expression of fat-1 gene in maligment cell enhances the occurring of cell apoptosis up to 35%～50%.Measurement of prostaglandin E2 found that the concentration of the PGE2 also markedly reduced.These data provide a foundation for applying ω-6 fatty acid desaturase gene as gene therapy of human breast cancer.更多还原