The functions of azurin of Pseudomonas aeruginosa and human mammaglobin-A on proapoptotic and cell cycle regulatory genes expression in the MCF-7 breast cancer cell line |
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Institution: | 1. Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran;2. Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran;3. Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, USA |
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Abstract: | Azurin protein of Pseudomonas aeruginosa is an anti-tumor agent against breast cancer and mammaglobin-A (MAM-A) protein is a specific antigen on the surface of MCF-7 for induction of cellular immune. The purpose of the present study was to investigate the effects of simultaneous expression of azurin and human MAM-A genes on the mRNA expression level of apoptosis-related and cell cycle genes in MCF-7 breast cancer cell line. The recombinant or empty plasmids were separately transferred into MCF-7 cells using Lipofectamine reagent. Flow cytometry was done to detect cell death and apoptosis. The expression of azurin and MAM-A genes were evaluated by IF assay, RT-PCR and western blot methods. Finally, apoptosis-related and cell cycle genes expression was examined in transformed and non-transformed MCF-7 cells by qPCR method. The successful expression of azurin and MAM-A genes in the MCF-7 cell were confirmed by RT-PCR, IF and western blotting. The apoptosis assay was showed a statistically significant (p < 0.05) difference after transfection. The expression of BAK, FAS, and BAX genes in transformed cells compare with non-transformed and transformed MCF-7 by pBudCE4.1 were increased statistically significant (p < 0.05) increases. Although, the increase of SURVIVIN and P53 expressions in transformed cells were not statistically significant (p > 0.05). Co-expression of azurin and MAM-A genes could induce apoptosis and necrosis in human MCF-7 breast cancer cells by up-regulation of BAK, FAS, and BAX genes. In future researches, it must be better the immune stimulation of pBudCE4.1-azurin-MAM-A recombinant vector in animal models and therapeutic approaches will be evaluated. |
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Keywords: | Azurin MAM-A MCF-7 Proapoptosis Cell cycle genes MCF-7"} {"#name":"keyword" "$":{"id":"k0040"} "$$":[{"#name":"text" "_":"Michigan Cancer Foundation-7 MAM-A"} {"#name":"keyword" "$":{"id":"k0050"} "$$":[{"#name":"text" "_":"mammaglobin-A IF"} {"#name":"keyword" "$":{"id":"k0070"} "$$":[{"#name":"text" "_":"Immunofluorescence q-PCR"} {"#name":"keyword" "$":{"id":"k0080"} "$$":[{"#name":"text" "_":"quantitative real-time PCR |
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