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

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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“…and Bacillus spp. [155]. Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, is widely used in ophthalmology, dermatology, and neurology [156].…”

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“…Among these, cupredoxins are metalloproteins involved in bacterial electron transport chain [8,9] showing different activities including antimalarial [10] and anticancer ones [5,[11][12][13][14][15]. The cupredoxin family includes azurins, rusticyanins, pseudoazurins, auracyanins, amicyanins, and halocyanins [16,17].…”