Abstract
Aim:
To construct a Bifidobacterium infantis/CD targeting gene therapy system and observe the antitumor effect of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system mediated by Bifidobacterium infantis on melanoma in vitro and in vivo.
Methods:
A recombinant CD/pGEX-1LamdaT plasmid was transfected into Bifidobacterium infantis by electroporation. Bifidobacterium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid was incubated with 5-FC anaerobically. Then the supernatant fluid was collected and added to melanoma B16-F10 cells to observe the killing effect for B16-F10 cells. Mice were inoculated with melanoma B16-F10 cells to establish animal models. The mice were then injected with 5-FC and Bifidobacterium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid.
Results:
Two segments of approximate 4.9 kb and 1.3 kb were extracted from the 6.2 kb recombinant plasmid, which were equal to the size of the pGEX-1LamdaT plasmid and CD gene, respectively. Sequencing results showed that the full length and sequence of nucleotide acid of the inserted gene in extracted recombinant plasmid was completely identical to the CD gene. In vitro, B16-F10 cells treated by supernatant fluid were remarkably damaged morphologically, and the cell growth was significantly inhibited. Experiments on the mice melanoma model showed that after treatment with a combination of transfected Bifidobacterium infantis and 5-FC, the tumor volume was significantly inhibited compared with controls.
Conclusion:
The foreign gene, CD gene, was correctly inserted into pGEX-1LambdaT plasmid and transferred into Bifidobacterium infantis. CD/5-FC suicide gene therapy system mediated by Bifidobacterium infantis demonstrated a good antitumor effect on melanoma in vitro and in vivo.
Similar content being viewed by others
Article PDF
References
Miller CR, Gustin AN, Buchsbaum DJ, Vickers SM, Manne U, Grizzle WE, et al. Quantitation of cytosine deaminase mRNA by real-time reverse transcription polymerase chain reaction: a sensitive method for assessing 5-fluorocytosine toxicity in vitro. Anal Biochem 2002; 30: 189–99.
Miyagi T, Koshida K, Hori O, Konaka H, Katoh H, Kitagawa Y, et al. Gene therapy for prostate cancer using the cytosine deaminase/uracil phosphoribosyltransferase suicide system. J Gene Med 2003; 5: 30–7.
Plumb JA, Bilsland A, Kakani R, Zhao J, Glasspool RM, Knox RJ, et al. Telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells to the pro-drug CB1954. Oncogene 2001; 20: 7797–803.
Brown NL, Lemoine NR . Clinical trials with GDEPT: cytosine deaminase and 5-fluorocytosine. Methods Mol Med 2004; 90: 451–7.
Xu G, McLeod HL . Strategies for enzyme/prodrug cancer therapy. Clin Cancer Res 2001; 7: 3314–24.
Denny WA . Prodrug strategies in cancer therapy. Eur J Med Chem 2001; 36: 577–95.
Liu SC, Minton NP, Giaccia AJ, Brown JM . Anticancer efficacy of systemically delivered anaerobic bacteria as gene therapy vectors targeting tumor hypoxia/necrosis. Gene Ther 2002; 9: 291–6.
Nuyts S, Van Mellaert L, Theys J, Landuyt W, Lambin P, Anne J . Clostridium spores for tumor-specific drug delivery. Anticancer Drugs 2002; 13: 115–25.
Yazawa K, Fujimori M, Amano J, Kano Y, Taniguchi S . Bifidobacterium longum as a delivery system for cancer gene therapy: selective localization and growth in hypoxic tumors. Cancer Gene Ther 2000; 7: 269–74.
Wu Y, Yi C, Wang S, Zhang M, Zhang J . Investigation on tumortargeting characteristics of Bifidobacterium infantis toward melanoma in mice. J Sichuan Univ 2003; 34: 435–8.
Vaupel PW . Oxygenation of solid tumors in drug resistance in oncology. In: Teicher BA, editor New York: Marcel Dekker; 1993, p 53–85.
Nagy H, Panis Y, Fabre M, Perrin H, Klatzmann D, Houssin D . Are hepatomas a good target for suicide gene therapy? An experimental study in rats using retroviral-mediated transfer of thymidine kinase gene. Surgery 1998; 123: 19–24.
Fujimori M, Amano J, Taniguchi S . The genus Bifidobacterium for cancer gene therapy. Curr Opin Drug Discov Devel 2002; 5: 200–3.
Yazawa K, Fujimori M, Nakamura T, Sasaki T, Amano J, Kano Y, et al. Bifidobacterium longum as a delivery system for gene therapy of chemically induced rat mammary tumors. Breast Cancer Res Treat 2001; 66: 165–70.
Nakamura T, Sasaki T, Fujimori M, Yazawa K, Kano Y, Amano J, et al. Cloned cytosine deaminase gene expression of Bifidobacterium longum and application to enzyme/pro-drug therapy of hypoxic solid tumors. Biosci Biotechnol Biochem 2002; 66: 2362–6.
Kim TB, Song SH, Kang SC, Oh DK . Quantitative comparison of lactose and glucose utilization in Bifidobacterium longum cultures. Biotechnol Prog 2003; 19: 672–5.
Makelainen H, Tahvonen R, Salminen S, Ouwehand AC . In vivo safety assessment of two Bifidobacterium longum strains. Microbiol Immunol 2003; 47: 911–4.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by CMB project (MERF-2002)
Rights and permissions
About this article
Cite this article
Yi, C., Huang, Y., Guo, Zy. et al. Antitumor effect of cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by Bifidobacterium infantis on melanoma. Acta Pharmacol Sin 26, 629–634 (2005). https://doi.org/10.1111/j.1745-7254.2005.00094.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2005.00094.x
Keywords
This article is cited by
-
Anti-cancer activity of human gastrointestinal bacteria
Medical Oncology (2022)
-
Pre-treatment with Bifidobacterium infantis and its specific antibodies enhance targeted radiosensitization in a murine model for lung cancer
Journal of Cancer Research and Clinical Oncology (2021)
-
Developing a new class of engineered live bacterial therapeutics to treat human diseases
Nature Communications (2020)
-
Engineering the gut microbiota to treat chronic diseases
Applied Microbiology and Biotechnology (2020)
-
Probiotic Bacteria: A Promising Tool in Cancer Prevention and Therapy
Current Microbiology (2019)