Construction of expression vector of siRNA specific for Alpha-fetoprotein and identification of its efficiency in FU97 gastric carcinoma cell line

Many studies have examined the biological activities of Alpha-fetoprotein (AFP); however, the modulatory role of AFP in neoplastic growth remains unclear. Three hairpin siRNA expressing plasmids (pSIREN -DNR -DsRed Express Donor - Vector - AFP - siRNA1/2/3) were constructed and transfected into FU97 (AFP - producing gastric adenocarcinoma cell line). At 48 h after transfection, the silencing efficiency to AFP gene expression was analyzed by RT - PCR and immunoassays. The influence to FU97 on its growth and apoptosis after silencing the AFP gene were observed by MTT and flow cytometry. The results of RT - PCR and immunoassays showed that the plasmids could inhibit AFP mRNA and protein expression at 48 h after transfection. The pSIREN - DNR - DsRed - Express Donor Vector - AFP - siRNA1 was more effective than other two. These plasmids could obviously inhibit FU97 cell growth and promote FU97 cell apoptosis.