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目的 :研究阳离子脂质体联合转铁蛋白(transferrin,Tf)介导甲胎蛋白 (AFP)启动子/增强子载体对分泌AFP的人肝癌细胞株的双重靶向作用。方法:测定比较阳离子脂质体Lipofectin、Lipofectin联合含双铁的人转铁蛋白[HTf(Fe)2]介导质粒 pDR2luc、pEBAFluc转染分泌AFP的人肝癌细胞株 (HepG2)、不分泌AFP的人肝癌细胞株 (SMMC7721)、人肾癌细胞株 (GRC)及非洲绿猴肾细胞 (COS7)的转染效率 ,转染效率以液体闪烁计数仪单光子计数法测定荧光素酶活性估计。结果 :(1)除COS7 外 ,其余3株细胞以Lipofectin +HTf(Fe)2介导pDR2luc的转染效率均比单用Lipofectin的转染效率高 (均P<0.01)。(2)以Lipofectin介导 pEBAFluc转染4株细胞 ,荧光素酶活性仅在能分泌AFP的HepG2 中表达。 (3)比较Lipofectin、Lipofectin +HTf(Fe) 2 介导 pEBAFluc转染HepG2 的效率 ,后者较前者高8倍多 (P<0.01)。结论 :Lipofectin +HTf(Fe)2介导含人AFP启动子/增强子的质粒pEBAFluc转染细胞时对分泌AFP的人肝癌细胞株有双重靶向作用
OBJECTIVE: To study the dual targeting of AFP-stimulated human hepatocellular carcinoma cell line by cationic liposome-mediated transferrin (Tf) -mediated AFP promoter / enhancer vector. METHODS: The human hepatoma cell line (HepG2) secreting AFP was transfected with Lipofectin and Lipofectin in combination with double ferro-human transferrin [HTf (Fe) 2] mediated plasmid pDR2luc and pEBAFluc. Transfection efficiency of human hepatocellular carcinoma cell line (SMMC7721), human renal cell carcinoma (GRC) and African green monkey kidney cells (COS7) were evaluated by luciferase activity assay by liquid scintillation counting single photon counting. Results: (1) The transfection efficiency of pDR2luc mediated by Lipofectin + HTf (Fe) 2 was higher than that transfected with Lipofectin alone (all P <0.01) except COS7. (2) Four cells were transfected with pEBAFluc by Lipofectin. The luciferase activity was only expressed in HepG2 secreting AFP. (3) Compared with Lipofectin, Lipofectin + HTf (Fe) 2 mediated the efficiency of pEBAFluc transfected HepG2, which was more than 8 times higher than the former (P <0.01). CONCLUSION: Lipofectin + HTf (Fe) 2 can double-target human hepatocarcinoma cell line secreting AFP when transfected with plasmid pEBAFluc containing AFP promoter / enhancer