Augmenting Anticancer Potential of Exotoxin A By Mutating Pseudomonas aeruginosa.
Yehia A. O. Ellazeik¹, Samah S. Abdelgawad², Essam MA Elsawy², Ahmed M. El-Wassef3.
¹Department of Botany, Faculty of Science, Mansoura University.
²Microbiology lab, Urology and Nephrology Center, Mansoura University.
3Department of Chemistry, Faculty of Science, Mansoura University.
Abstract: Cancer is a group of more than 100 different and distinctive types of diseases. It is an abnormal growth of cells which tend to proliferate in an uncontrolled manner forming masses and in some cases dislodge and spread all over the body. Luckily, a number of bacteria including Pseudomonas aeruginosa produce some virulence factors that help in combating cancer such as exotoxin A (ETA). This toxin arrests protein synthesis and induces apoptosis in cancer cells. During this study 68 Pseudomonades were isolated from urine samples collected from patients with urinary tract infection during the period Sept. 2009- Feb. 2010. Classical bacteriological, molecular and automated methods were used to identify all them. Based on the protein banding patterns, the 68 isolates were re-grouped to 30according to the results of Sodium dedocylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), from these 30 isolates only three gave the typical PCR product for Exo A gene using its specific primers. ETA production and toxicity were enhanced by mutating the three wild-type isolates. The crude and partially purified ETA from the selected three wild-type isolates of Pseudomonas aeroginosa (1, 8, 15) and two UV mutants (3-1 and 16-15) showed a promising inhibitory activity against the MCF-7 cell line of breast carcinoma. The IC50 (inhibition concentration) of the five organisms were 14.1, 35.6, 36.8, 5.3, 3.4µg, respectively. The mutation increased the anticancer activity of ETA 2-fold for one mutant and 10-fold to the second mutant. No change in the molecular weight of the mutated protein was found and the exact nature of its anticancer activity is under further investigation.
[Yehia A. Ellazeik, Samah S. Abdelgawad,Essam MA Elsawy and Ahmed M. El-Wassef. Augmenting Anticancer Potential of Exotoxin A By Mutating Pseudomonas aeruginosa. J Am Sci 2013;9(3):312-321]. (ISSN: 1545-1003). http://www.jofamericanscience.org. 55
Key Words: Pseudomonas aeruginosa;virulence factors;cancer;Exotoxin A(ETA);PCR. Full Text 55