摘要
目的:构建携铜绿假单胞菌外毒素PE38基因及甲胎蛋白(AFP)启动子的重组载体,并研究其在体外对AFP阳性肝细胞癌(hepatocellularcarcinoma,HCC)的靶向杀伤作用。方法:构建重组免疫毒素表达质粒pAFP-PE38,通过转染细胞观察其作用,RT-PCR法测定PE38 mRNA表达,CCK-8法检测细胞毒性。结果:各组细胞转染质粒pAFP-PE38后仅AFP阳性的HepG2细胞表达PE38 mRNA,且形态发生改变,生长显著受抑(P<0.01),48 h和72 h抑制率分别为27.2%、58.3%,而AFP阴性的PC-3和HeLa细胞未受明显影响。结论:PE38基因真核表达载体可实现HCC基因治疗的靶向性和高效性,有望成为肝细胞癌靶向基因治疗的有力工具。
AIM：To construct a gene-modified hepatocellular carcinoma（HCC）specific PE38 expression vector regulated by cis-acting element of AFP,and explore its anti-HCC effect in vitro.METHODS：Eukaryotic expression plasmid pAFP-PE38 was constructed and then transfected into different cell lines.The expression level of PE38 mRNA was detected by RTPCR. Cytotoxicity was detected by cell counting kit-8（CCK-8）.RESULTS：PE38 mRNA was detected only in AFP-positive HepG2 cells after plasmid transfection.Significant morphological changes and growth inhibition（P〈0.01） were observed in HepG2 cells,the inhibition rate at 48 h and 72 h were 27.2%and 58.3%,respectively; whereas no such changes were found in AFP-negative PC-3 cells and HeLa cells.CONCLUSION： PE38 gene eukaryotic expression vector can selectively targets on HCC cells with high efficacy,which might be used as an effective tool for HCC gene therapy.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2010年第4期372-375,共4页
Chinese Journal of Clinical Pharmacology and Therapeutics
