[Detection and comparison of transcriptional activities of tumor-specific survivin and alpha-fetoprotein promoters in human hepatocellular carcinoma cells].
OBJECTIVE
To 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.


METHODS
The 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.


RESULTS
Both 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.


CONCLUSIONS
Our 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.
Kuang
Ge
G
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Huang
Ai-long
AL
Tang
Ni
N
D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D018390:Gene Targeting; D015316:Genetic Therapy; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008113:Liver Neoplasms / Q000235:genetics*; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D000077022:Survivin; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000235:genetics*
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; D000509:alpha-Fetoproteins