Volume 71, Issue 11 (February 2014)
Tehran Univ Med J 2014, 71(11): 691-699 |
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Ghaderi N, Esazadeh K, Shoae Hasani A. Utilization of Lambda bacteriophage as an Apoptin effective delivery platform to the BT-474 human breast carcinoma. Tehran Univ Med J 2014; 71 (11) :691-699
URL: http://tumj.tums.ac.ir/article-1-5783-en.html
URL: http://tumj.tums.ac.ir/article-1-5783-en.html
1- Department of Microbiology, Faculty of Science, Islamic Azad University Lahijan Branch, Gilan, Iran. , narminghaderi@yahoo.com
2- Department of Microbiology, Faculty of Science, Islamic Azad University Lahijan Branch, Gilan, Iran.
3- Department of Stem Cell and Tissue Engineering, Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
2- Department of Microbiology, Faculty of Science, Islamic Azad University Lahijan Branch, Gilan, Iran.
3- Department of Stem Cell and Tissue Engineering, Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Abstract: (8634 Views)
Background: Apoptin is a protein from chicken anemia virus that could induce apoptosis specifically in the cancer cells but it has not any effect in the normal cells. Phage therapy is a novel field of cancer therapy and phage nanobioparticles (NBPs) such as λ phage could be modified to deliver and express genetic cassettes into eukaryotic cells safely in contrast with animal viruses. The bacteriophages like Lambda could be manipulated to deliver genetic cassettes into eukaryotic cells and express the gene safely. We developed the safe way for the expression of Apoptin gene via Lambda bacteriophage in the human tumors.
Methods: At first the Apoptin clone was produced and then transferred into ZAP-CMV plasmid through BamH-I and HinD-III restriction sites. Then this construct inserted into the Lambda phage in the Escherichia coli host cell. The expression of Apoptin in the recombinant construct was evaluated via RT-PCR and Western Blot analysis. The anti tumor function of expressed protein was measured in the BT-474 cells that was hosted by nude mice.
Results: Transfection of breast carcinoma cells by Lambda bacteriophage containing λZAP-Apoptin-CMV was inhibited the tumor growth significantly but did not any effect on normal cells. The expression of this protein was very high in tumor cells and prevented the death of tumor bearing nude mice. The penetration and spreading of Apoptin construct by bacteriophage Lambda was significantly high but the Apoptin plasmid had very little expression in BT-474 cell, directly. Transfection with NBPs carrying λZAP-CMV-Apoptin significantly inhibited growth of all the breast carcinoma cell lines in vitro, but had no effect on normal cells.
Conclusion: Utilization of recombinant Lambda bacteriophage as a safe expression vector has been confirmed. Apoptin was induced apoptosis specifically in the tumors in vivo. Use of such construct is a very safe way to treat cancer in human. The results presented here reveal important features of λ nanobioparticles to serve as safe delivery and expression platform for human cancer therapy.
Type of Study: Original Article |
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