癌胚抗原肿瘤基因疫苗胶囊的研制

Preparation of carcinoembryonic antigen positive tumor gene vaccine capsules for oral use

目的研制癌胚抗原(CEA)阳性肿瘤基因疫苗结肠溶胶囊。方法通过分子克隆技术将加强型绿色荧光EGFP cDNA连接于CEA cDNA的上游,构建示踪子EGFP-CEA融合基因疫苗pVGC2。使用特制的微小结肠溶空心胶囊制备鼠用基因疫苗口服胶囊。收集小鼠口服用药后的腹腔游离细胞,采用荧光显微镜观察绿色荧光以便评估EGFP-CEA融合蛋白质的表达情况。结果所构建的重组质粒的目的基因片段测序结果证实所接入的EGFP-CEA cDNA序列与母本cDNA序列100%一致。所制备的口服胶囊的平均溶出度为(91.1±3.65)%。检测结果显示,口服胶囊后小鼠腹腔游离细胞中可见到绿色荧光。结论成功地制备了口服基因疫苗胶囊。小鼠腹腔游离细胞中绿色荧光融合蛋白的表达预示,口服基因疫苗胶囊是一种潜在可行的抗肿瘤免疫途径。

OBJECTIVE To prepare and study the oral carcinoembryonic antigen （CEA） positive tumor gene vaccine capsules.METHODS By using a cloning method, a EGFP-CEA cDNA fusion gene vaccine pVGC2 was constructed as a tracer in which EGFP cDNA fragment was linked to the upstream of CEA cDNA. With a kind of subminiature hollow capsules which dissolved at the colon site, the oral gene vaccine capsules were prepared for mice. The expression of the protein of the ECFP-CEA gene in mice was estimated after the capsules were administered orally. The green fluorescent features in the free cells collected from the abdominal cavity were observed by fluoromi-croscope. RESULTS The nucleotide sequences of the target gene fragments of the recombinant plasmid were verified that the acquired EGFP cDNA sequence was coincidence with the original cDNA（ 100% ） provided. The average dissolution of the oral capsules was （91.1±3.65）% . And the results showed that the free cells had a visible green fluorescence after the oral treatment. CONCLUSION A kind of oral gene vaccine capsules were successfully prepared. And the green fluorescent expression of the fusion protein of the gene vaccine in the cells indicated the oral medicine path of the capsules was a potentially immune way for anticancer.