阳离子脂质体联合转铁蛋白介导肝癌组织特异性载体在人肝癌细胞株中的表达

Expression of Human Liver Cancer Specific Carrier in Human Hepatocarcinoma Cell Line Mediated by Cationic Liposome Combined with Transferrin

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)

Objective:To study the double-targeting effect onα-fetoprotein(AFP)-positive human hepatocarcinoma cell line of the liposome plus transferrin(TF)mediated transfection of tissue-specific vector.Methods:Plasmid pDR 2 luc and pEBAFluc contained the firefly luciferase reporter gene under the control of RSV-LTR promoter and the AFP promoter respectively.Four cell lines including human hepatocarcinoma cell line HepG 2 [TfR(+),AFP(+)],SMMC7721[TfR(+),AFP(-)],human renal carcinoma cell line GRC[TfR(+),AFP(-)],and simian kidney cell line COS 7 [TfR(+),AFP(-)] were used to be transfected by cationic liposome Lipofectin and Lipofectin plus human transferrin with two molecules of Fe 3+ [HTf(Fe) 2 ] respectively.The transfection efficiency was evaluated by the activity of expressed luciferase.Results:(1)Comparing the Lipofectin-HTf(Fe) 2 PDR 2 luc complexes with the Lipfectin-pDR 2 luc complexes,the luciferase activity was at least8times higher in HepG 2 and SMMC7721(P<0.01),and at least4times higher in GRC(P<0.01).(2)When transfected with Lipofectin-pEBAFluc,luciferase activity only expressed in HepG 2 ,and did not express in other three cell lines.(3)The luciferase activity was at least8times higher in HepG 2 transfected with Lipofectin-pEBAFluc-HTf(Fe) 2 complexes.Conclusion:Lipofectin plus HTf(Fe) 2 mediated pEBAFluc delivery has double targeting effects on the AFP-positive hepatocarcinoma cells.