Adenovirus-Based p53 Gene Therapy in Ovarian Cancer

JOSEPH T. SANTOSO, DE CHU TANG, STEVEN B. LANE, JACLYN HUNG, DEBORAH J. REED, CAROLYN Y. MULLER, DAVID P. CARBONE, JOSEPH A. LUCCIIII, DAVID SCOTTMILLER, MICHAEL J. MATHIS

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Abstract

Mutations of the p53 tumor suppressor gene are the most common molecular genetic abnormality to be described in ovarian cancer. To determine the feasibility of mutant p53 as a molecular target for gene therapy in ovarian cancer, we constructed an adenovirus vector containing the wild-type p53 gene. The ability of this adenovirus construct (Ad-CMV-p53) to express p53 protein was examined by Western blot analysis in the H358 lung cancer cell line, which has a homozygous deletion of the p53 gene. The ability of the adenovirus vector system to infect ovarian cancer cells was tested using an adenovirus containing the β-galactosidase reporter gene under the control of the CMV promoter (Ad-CMV-βgal). The ovarian cancer cell line 2774, which contains an Arg273His p53 mutation, was infected with Ad-CMV-βgal, and the infected cells were assayed for β-galactosidase activity after 24 hr. To test the ability of wild-type p53 to inhibit cell growth, the 2774 cell line was infected with Ad-CMV-p53 or Ad-CMV-βgal, and the effect of these agents on the growth of 2774 cells was determined using anin vitrogrowth inhibition assay. Western blot analysis of lysates from H358 cells infected with Ad-CMV-p53 showed expression of wild-type p53 protein. When 2774 cells were infected with Ad-CMV-βgal at a multiplicity of infection (m.o.i.) of 10 PFU/cell, >90% of cells showed β-galactosidase activity, demonstrating that these cells are capable of efficient infection by the adenovirus vector. Growth of 2774 cells infected with Ad-CMV-p53 was inhibited by >90% compared to noninfected cells. The ability of the adenovirus vector to mediate high-level expression of infected genes and the inhibitory effect of Ad-CMV-p53 on the 2774 cell line suggests that the Ad-CMV-p53 could be further developed into a therapeutic agent for ovarian cancer.

Original languageEnglish (US)
Pages (from-to)171-178
Number of pages8
JournalGynecologic Oncology
Volume59
Issue number2
DOIs
StatePublished - Nov 1995

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p53 Genes
Adenoviridae
Genetic Therapy
Ovarian Neoplasms
Galactosidases
Cell Line
Growth
Western Blotting
Adenoviridae Infections
Mutation
Tumor Suppressor Genes
Reporter Genes
Molecular Biology
Lung Neoplasms
Proteins
Gene Expression

ASJC Scopus subject areas

  • Oncology
  • Obstetrics and Gynecology

Cite this

SANTOSO, JOSEPH. T., TANG, DE. CHU., LANE, STEVEN. B., HUNG, JACLYN., REED, DEBORAH. J., MULLER, CAROLYN. Y., ... MATHIS, MICHAEL. J. (1995). Adenovirus-Based p53 Gene Therapy in Ovarian Cancer. Gynecologic Oncology, 59(2), 171-178. https://doi.org/10.1006/gyno.1995.0002

Adenovirus-Based p53 Gene Therapy in Ovarian Cancer. / SANTOSO, JOSEPH T.; TANG, DE CHU; LANE, STEVEN B.; HUNG, JACLYN; REED, DEBORAH J.; MULLER, CAROLYN Y.; CARBONE, DAVID P.; LUCCIIII, JOSEPH A.; SCOTTMILLER, DAVID; MATHIS, MICHAEL J.

In: Gynecologic Oncology, Vol. 59, No. 2, 11.1995, p. 171-178.

Research output: Contribution to journalArticle

SANTOSO, JOSEPHT, TANG, DECHU, LANE, STEVENB, HUNG, JACLYN, REED, DEBORAHJ, MULLER, CAROLYNY, CARBONE, DAVIDP, LUCCIIII, JOSEPHA, SCOTTMILLER, DAVID & MATHIS, MICHAELJ 1995, 'Adenovirus-Based p53 Gene Therapy in Ovarian Cancer', Gynecologic Oncology, vol. 59, no. 2, pp. 171-178. https://doi.org/10.1006/gyno.1995.0002
SANTOSO JOSEPHT, TANG DECHU, LANE STEVENB, HUNG JACLYN, REED DEBORAHJ, MULLER CAROLYNY et al. Adenovirus-Based p53 Gene Therapy in Ovarian Cancer. Gynecologic Oncology. 1995 Nov;59(2):171-178. https://doi.org/10.1006/gyno.1995.0002
SANTOSO, JOSEPH T. ; TANG, DE CHU ; LANE, STEVEN B. ; HUNG, JACLYN ; REED, DEBORAH J. ; MULLER, CAROLYN Y. ; CARBONE, DAVID P. ; LUCCIIII, JOSEPH A. ; SCOTTMILLER, DAVID ; MATHIS, MICHAEL J. / Adenovirus-Based p53 Gene Therapy in Ovarian Cancer. In: Gynecologic Oncology. 1995 ; Vol. 59, No. 2. pp. 171-178.
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abstract = "Mutations of the p53 tumor suppressor gene are the most common molecular genetic abnormality to be described in ovarian cancer. To determine the feasibility of mutant p53 as a molecular target for gene therapy in ovarian cancer, we constructed an adenovirus vector containing the wild-type p53 gene. The ability of this adenovirus construct (Ad-CMV-p53) to express p53 protein was examined by Western blot analysis in the H358 lung cancer cell line, which has a homozygous deletion of the p53 gene. The ability of the adenovirus vector system to infect ovarian cancer cells was tested using an adenovirus containing the β-galactosidase reporter gene under the control of the CMV promoter (Ad-CMV-βgal). The ovarian cancer cell line 2774, which contains an Arg273His p53 mutation, was infected with Ad-CMV-βgal, and the infected cells were assayed for β-galactosidase activity after 24 hr. To test the ability of wild-type p53 to inhibit cell growth, the 2774 cell line was infected with Ad-CMV-p53 or Ad-CMV-βgal, and the effect of these agents on the growth of 2774 cells was determined using anin vitrogrowth inhibition assay. Western blot analysis of lysates from H358 cells infected with Ad-CMV-p53 showed expression of wild-type p53 protein. When 2774 cells were infected with Ad-CMV-βgal at a multiplicity of infection (m.o.i.) of 10 PFU/cell, >90{\%} of cells showed β-galactosidase activity, demonstrating that these cells are capable of efficient infection by the adenovirus vector. Growth of 2774 cells infected with Ad-CMV-p53 was inhibited by >90{\%} compared to noninfected cells. The ability of the adenovirus vector to mediate high-level expression of infected genes and the inhibitory effect of Ad-CMV-p53 on the 2774 cell line suggests that the Ad-CMV-p53 could be further developed into a therapeutic agent for ovarian cancer.",
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