Construction Of Expression Vector Of SiRNA Specific For Alpha-fetoprotein And Identification Of Its Efficiency In FU97 Gastric Carcinoma Cell Line

Objective Since alpha-fetoprotein(AFP) was found, as amarker for fetal disease and some tumors , AFP plays an important role in the areas of clinic diagnosis and disease detection. A lot of researches in its clinic significance have been done. However, in the past several decades, the role of AFP in the regulation of cancer growth has received little attention. Only during the last decade has the modulatory role of AFP in neoplastic growth been realized and implemented in experimental models. Here we take an AFP-producing gastric adenocarcinoma cell line, FU97, as a study aim. By the technology of RNA interfering, we want to construct three different siRNA expression vectors (pDonor Vector AFP siRNA1/2/3) specific for human AFP gene , then to observe their silencing effect on AFP gene in FU97 as well as to study the influence to tumor cell on the growth and apopotosis after silencing the AFP gene. To find the function of AFP in the growth and development of AFP related tumors and discuss the potential applications of AFP in the gene therapy of AFP related tumors. Methods To choose three qualified target sequences in AFP gene , there are: â‘ CAGGGAGACATTCATGAAC, begin at 508 point, the GC content of the selected 19-base oligonucleotide sequence is 47 % ; â‘¡ CTGGAACGTGGTCAATGTA, begin at 968 point, the GC content is 47%; â‘¢...