Evaluation of tumor-specific promoter activities in melanoma

B. Lu, Sharmila K. Makhija, D. M. Nettelbeck, A. A. Rivera, M. Wang, S. Komarova, F. Zhou, M. Yamamoto, H. J. Haisma, R. D. Alvarez, D. T. Curiel, Z. B. Zhu

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

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), α-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.

Original languageEnglish (US)
Pages (from-to)330-338
Number of pages9
JournalGene Therapy
Volume12
Issue number4
DOIs
StatePublished - Feb 2005
Externally publishedYes

Fingerprint

Carcinogens
Melanoma
Luciferases
Genetic Therapy
Glycoproteins
Primary Cell Culture
Cyclooxygenase 2
Cell Line
CXCR4 Receptors
Poly A
Neoplasm Genes
Liver
Melanocytes
Tumor Cell Line
Transgenes
Reporter Genes
Chemokines
Messenger RNA

ASJC Scopus subject areas

  • Genetics

Cite this

Lu, B., Makhija, S. K., Nettelbeck, D. M., Rivera, A. A., Wang, M., Komarova, S., ... Zhu, Z. B. (2005). Evaluation of tumor-specific promoter activities in melanoma. Gene Therapy, 12(4), 330-338. https://doi.org/10.1038/sj.gt.3302385

Evaluation of tumor-specific promoter activities in melanoma. / Lu, B.; Makhija, Sharmila K.; Nettelbeck, D. M.; Rivera, A. A.; Wang, M.; Komarova, S.; Zhou, F.; Yamamoto, M.; Haisma, H. J.; Alvarez, R. D.; Curiel, D. T.; Zhu, Z. B.

In: Gene Therapy, Vol. 12, No. 4, 02.2005, p. 330-338.

Research output: Contribution to journalArticle

Lu, B, Makhija, SK, Nettelbeck, DM, Rivera, AA, Wang, M, Komarova, S, Zhou, F, Yamamoto, M, Haisma, HJ, Alvarez, RD, Curiel, DT & Zhu, ZB 2005, 'Evaluation of tumor-specific promoter activities in melanoma', Gene Therapy, vol. 12, no. 4, pp. 330-338. https://doi.org/10.1038/sj.gt.3302385
Lu B, Makhija SK, Nettelbeck DM, Rivera AA, Wang M, Komarova S et al. Evaluation of tumor-specific promoter activities in melanoma. Gene Therapy. 2005 Feb;12(4):330-338. https://doi.org/10.1038/sj.gt.3302385
Lu, B. ; Makhija, Sharmila K. ; Nettelbeck, D. M. ; Rivera, A. A. ; Wang, M. ; Komarova, S. ; Zhou, F. ; Yamamoto, M. ; Haisma, H. J. ; Alvarez, R. D. ; Curiel, D. T. ; Zhu, Z. B. / Evaluation of tumor-specific promoter activities in melanoma. In: Gene Therapy. 2005 ; Vol. 12, No. 4. pp. 330-338.
@article{29f7926ae25d451e868396fa46a6b127,
title = "Evaluation of tumor-specific promoter activities in melanoma",
abstract = "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), α-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.",
author = "B. Lu and Makhija, {Sharmila K.} and Nettelbeck, {D. M.} and Rivera, {A. A.} and M. Wang and S. Komarova and F. Zhou and M. Yamamoto and Haisma, {H. J.} and Alvarez, {R. D.} and Curiel, {D. T.} and Zhu, {Z. B.}",
year = "2005",
month = "2",
doi = "10.1038/sj.gt.3302385",
language = "English (US)",
volume = "12",
pages = "330--338",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Evaluation of tumor-specific promoter activities in melanoma

AU - Lu, B.

AU - Makhija, Sharmila K.

AU - Nettelbeck, D. M.

AU - Rivera, A. A.

AU - Wang, M.

AU - Komarova, S.

AU - Zhou, F.

AU - Yamamoto, M.

AU - Haisma, H. J.

AU - Alvarez, R. D.

AU - Curiel, D. T.

AU - Zhu, Z. B.

PY - 2005/2

Y1 - 2005/2

N2 - 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), α-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.

AB - 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), α-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.

UR - http://www.scopus.com/inward/record.url?scp=20044391056&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20044391056&partnerID=8YFLogxK

U2 - 10.1038/sj.gt.3302385

DO - 10.1038/sj.gt.3302385

M3 - Article

VL - 12

SP - 330

EP - 338

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 4

ER -