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构建由癌胚抗原 (CEA)启动子控制报道基因增强型绿色荧光蛋白 (EGFP)表达的重组表达质粒pCEA EGFP .用转染细胞后检测荧光的方法对CEA阳性细胞进行简便、直观的检测 ,并结合流式细胞计数对CEA启动子在人结直肠腺癌细胞LS 1 74T、结肠癌细胞SW 4 80、肺腺癌细胞A5 49、人宫颈癌细胞HeLa和人喉癌细胞HEp 2中的活性进行了分析 ,发现其在SW4 80、LS 1 74T、A5 49中活性较强 ,而在HeLa和HEp 2中无活性 .构建由CEA启动子控制凋亡基因bak表达的重组表达质粒pCEA bak ,转染HeLa及SW 4 80细胞 ,用Hoechst332 5 8染色及PI染色 流式细胞计数分析的方法证明 ,pCEA bak转染能够特异性引起SW 4 80细胞的凋亡 .结果表明 ,CEA启动子具有很好的特异性 ,CEA介导bak基因的方法可望用于CEA阳性癌细胞的靶向性基因治疗 .
To construct a recombinant plasmid pCEA EGFP whose expression of enhanced green fluorescent protein (EGFP) was controlled by the promoter of CEA promoter, the CEA positive cells were detected by fluorescence detection after transfected cells, The activity of the CEA promoter in human colorectal adenocarcinoma cells LS 1 74T, colon cancer cells SW 4 80, lung adenocarcinoma cells A549, human cervical carcinoma cells HeLa and human laryngeal carcinoma cells HEp 2 was analyzed by flow cytometry , It was found to be highly active in SW480, LS1 74T, A549 and inactive in HeLa and HEp 2. The recombinant expression plasmid pCEAbak, in which the CEA promoter controls the apoptosis gene bak, was constructed and transfected HeLa and SW480 cells, using Hoechst3328 staining and PI staining flow cytometry analysis showed that, pCEA bak transfection can specifically cause apoptosis in SW480 cells.The results show that the CEA promoter has a good Specific, CEA-mediated approach to the bak gene is expected to be used for targeted gene therapy in CEA-positive cancer cells.