Evaluation of tumor-specific promoter activities in melanoma.
Gene therapy is a novel therapy for melanoma. To date, however, there is still no powerful tumor specific promoter (TSP) to restrict the transgene expression in melanoma cells. In order to define a useful TSP for targeting in the context of melanoma gene therapy, four promoters, the cyclooxygenase-2 (Cox-2), alpha-chemokine SDF-1 receptor (CXCR4), epithelial glycoprotein 2 (EGP-2), and survivin, were tested in both established melanoma cell lines and primary melanoma cells. We employed recombinant adenoviral vectors (reAds) each with a candidate TSP (the Cox-2, CXCR4, EGP-2, or survivin), a reporter luciferase gene, and a poly-A signal, all of which were inserted into the E1-deleted region. A reAdGL3Bcytomegalovirus (CMV), containing the CMV promoter and luciferase gene, was used as a positive control to normalize the luciferase activity. Luciferase activity was measured in multiple tumor cell lines and two primary melanoma cell cultures after infection with reAds. Human epithelial melanocytes, HEM, were used as normal control. In contrast to three other promoters, the survivin promoter exhibited the highest activities within both melanoma cell lines and primary melanoma cells, but not in HEMs. Additionally, the survivin promoter exhibited very low activities in major mouse organs including the liver, in vivo. EGP-2 is not active in melanoma; messenger RNA expressions were correlated to promoter activities both in melanoma cell lines and primary cell cultures. Thus, these data suggest that the survivin promoter achieved a 'tumor-on/liver-off' profile, and thus represents a potentially useful tumor-specific promoter with applications for transcriptional targeting of Ad vector-based cancer gene therapy or oncolysis to melanoma.
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B
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Department of Medicine, Division of Human Gene Therapy, The Gene Therapy Center, 901 19th Street S., University of Alabama at Birmingham, Birmingham, AL 35291, USA.
Makhija
S K
SK
Nettelbeck
D M
DM
Rivera
A A
AA
Wang
M
M
Komarova
S
S
Zhou
F
F
Yamamoto
M
M
Haisma
H J
HJ
Alvarez
R D
RD
Curiel
D T
DT
Zhu
Z B
ZB
D000256:Adenoviridae / Q000235:genetics; D000954:Antigens, Surface / Q000235:genetics; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2; D000071858:Epithelial Cell Adhesion Molecule; D015870:Gene Expression; D018390:Gene Targeting; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008545:Melanoma / Q000628:therapy*; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009363:Neoplasm Proteins; D011401:Promoter Regions, Genetic; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics; D019718:Receptors, CXCR4 / Q000235:genetics; D012878:Skin Neoplasms / Q000628:therapy*; D000077022:Survivin; D014161:Transduction, Genetic / Q000379:methods; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; C107489:BIRC5 protein, human; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D051034:Inhibitor of Apoptosis Proteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D019718:Receptors, CXCR4; D000077022:Survivin; D008156:Luciferases; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases