OBJECTIVETo detect and compare the transcriptional activities of tumor-specific survivin and AFP promoters in various hepatocellular carcinoma cell lines and to lay some groundwork for targeting gene therapy in human hepatocellular carcinoma.

METHODSThe fragment of survivin and AFP promoters were acquired by PCR amplification and were cloned into the reporter plasmid pGL3-Basic, which contained a luciferase gene. The constructed eukaryotic expression plasmid pGL3-SUR and pGL3-AFP, in which the expression of the luciferase was derived by survivin or the AFP promoter, were transfected into three HCC cell lines. At 24 hours post transfection (p.t.), the activity of the luciferase was determined with Dual-Luciferase Reporter Assay System. A pGL3-CMV, containing the CMV promoter controlled luciferase gene, was used as a positive control.

RESULTSBoth survivin and AFP promoters had transcriptional activities in all three HCC cell lines and the transcriptional activity of the survivin promoter was much higher than the AFP promoter (52-98 times) and reached a level of 16% approximately 21% of the transcriptional activity of the CMV promoter.

CONCLUSIONOur data reveals that the survivin promoter possesses a high transcriptional activity in all three established HCC cell lines and may serve as a useful tool for transcriptional targeting gene therapy of HCCs.