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5-fluorouracil and hydroxyurea enhance adenovirus-mediated transgene expression in colon and hepatocellular carcinoma cells

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Abstract

Purpose

To investigate the efficient transduction of tumor cells which remains a major limitation of cancer gene therapy..

Methods

In this study, we tested whether treatment with antimetabolic drugs 5-FU and hydroxyurea (HU) could improve adenovirus-mediated gene expression in tumor cells.

Results

We found that 5-FU and HU treatment significantly increased β-gal activity in adenovirus (Ad.CMVBG)-infected human colon carcinoma (LoVo) and hepatocellular carcinoma (SMMC7721) cells in a dose- and time-dependent manner. These increases were maximized at 5.01±0.42-fold and 3.32±0.32-fold for 5-FU (50 μM), and at 6.60±0.50-fold and 4.82±0.43-fold for HU (5 mM) treatment, respectively, after 48 h infection. Transient increases in viral uptake, determined by real-time PCR for viral DNA content and by confocal microscopy for viral particles, were observed in 5-FU or HU-treated cells that partially contribute to the overall increases of β-gal expression. Moreover, mRNA levels for the β-gal gene in infected cells were significantly increased in both LoVo and SMMC7721 cells by 5-FU and HU treatment in contrast to the inhibition of viral DNA replication and the unchanged mRNA levels for α-actin gene. The induction appeared to be the result of enhanced transcription since β-gal mRNA half-life was not affected by drug treatment. However, similar induction was not detected in CMV-β-gal-expressing stable cells, suggesting that an adenovirus-associated mechanism might be involved in this induction.

Conclusions

Our findings suggest that it may be possible to improve tumor cell transduction by adenovirus using chemotherapy.

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Abbreviations

yCD:

Yeast cytosine deaminase

5-FC:

5-fluorocytosine

5-FU:

5-fluorouracil

HU:

Hydroxyurea

β-gal:

β-galactosidase

CMV:

Cytomegalovirus

Ad.CMVBG:

E1-deleted adenovirus containing β-gal gene

PCR:

Polymerase chain reaction

CAR:

Coxsackie-adenovirus receptor

pfu:

Plaque-forming units

MOI:

Multiplicity of infection

RT-PCR:

Reverse transcription-PCR

DRB:

5,6-dichlorobenzimidazole

FACS:

Fluorescence-activated cell sorting

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Authors and Affiliations

  1. Department of Radiation Oncology, Room 3111, University of Michigan, 1331 E Ann Street, Ann Arbor, MI , 48109-0582, USA

    Xiao W. Huang, Theodore S. Lawrence & Ming Zhang

  2. Liver Cancer Institute & Zhongshan Hospital, Fudan University, Shanghai, P.R. China

    Xiao W. Huang & Zhao Y. Tang

Authors
  1. Xiao W. Huang
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  2. Zhao Y. Tang
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  3. Theodore S. Lawrence
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  4. Ming Zhang
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Corresponding author

Correspondence to Ming Zhang.

Additional information

Supported by NIH grants CA80145, CA84117, Cancer Center Core Grant CA46592, and a Development and Feasibility Grant from the University of Michigan Center for Gene Therapy.

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Huang, X.W., Tang, Z.Y., Lawrence, T.S. et al. 5-fluorouracil and hydroxyurea enhance adenovirus-mediated transgene expression in colon and hepatocellular carcinoma cells. J Cancer Res Clin Oncol 131, 184–190 (2005). https://doi.org/10.1007/s00432-004-0608-1

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  • DOI: https://doi.org/10.1007/s00432-004-0608-1

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