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Background and Objective: Cervical cancer is the fourth most common cancer in the world. The most important issue in cancer treatment is the destruction of cancer cells in the presence of normal cells. For this reason, it is necessary to use natural resources such as plants to treat cancer. The aim of this study is to evaluate the antibacterial effects of the ethanolic extract of Alhagi maurorum on Staphylococcus aureus and Pseudomonas aeruginosa and cytotoxicity on HeLa cervical cancer cell line. Methods: In this experimental study, first the ethanolic extract of Alhagi maurorum was prepared, and then the two standard strains of Staphylococcus aureus (ATCC: 25923) and Pseudomonas aeruginosa (ATCC: 27853) were lyophilized by culturing in nutrient medium. In order to confirm the standard strains, biochemical tests were performed. Microdilution method was used to determine the minimum inhibitory concentration and after obtaining the minimum inhibitory concentration, the minimum bactericidal concentration was evaluated. Furthermore, the effects of the cytotoxicity of the extract at concentrations of 0.1, 10, 50, 100, 500 and 1000 μg/ml was evaluated on the HeLa cell line in a period of 48 hours using the MTT method and comparing its toxicity with the cisplatin group (positive control group). Findings: Ethanol extract of Alhagi maurorum at a concentration of 50 μg/ml reduced the growth of cancer cells, and in the statistical comparison, 50, 500 and 1000 μg concentrations revealed significant differences (p<0.05). According to minimum inhibitory concentration results, the minimum growth inhibitory concentration of the extract on the standard strain of Staphylococcus aureus and Pseudomonas aeruginosa was reported to be 4000 and 16000 μg/ml, respectively, and according to minimum bactericidal concentration results, the minimum bactericidal concentration of the extract was found 4 times the minimum inhibitory concentration (16000 μg/ml) in Staphylococcus aureus (ATCC: 25923), but it was not lethal in Pseudomonas aeruginosa (ATCC: 27853). Conclusion: The results of the study showed that the ethanolic extract of Alhagi maurorum affected HeLa cells through antioxidant activity and inhibited their growth, and according to minimum inhibitory concentration and minimum bactericidal concentration results, it was also shown that the most inhibitory effect was on the standard strain of Staphylococcus aureus while it showed no effects on the strain of Pseudomonas aeruginosa. [ABSTRACT FROM AUTHOR]