Volume 8, Issue 3 (Aug 2020)
Res Mol Med (RMM) 2020, 8(3): 107-116 |
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Khalafi A, Moradian F, Rafiei A. Anticancer Effect of Bovine Lactoferrin on Breast Cancer Cell Line MCF7 and the Evaluation of Bax and Bak Genes Expression Involved in Apoptosis. Res Mol Med (RMM) 2020; 8 (3) :107-116
URL: http://rmm.mazums.ac.ir/article-1-372-en.html
URL: http://rmm.mazums.ac.ir/article-1-372-en.html
1- Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran. , amir_khalafi1998@yahoo.com
2- Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.
3- Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
2- Department of Basic Sciences, Sari Agricultural Sciences and Natural Resources University, Sari, Iran.
3- Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
Abstract: (2626 Views)
Background: Lactoferrin is a glycoprotein with antimicrobial, antioxidant, immune-modulating, antiviral, and most importantly anticancer properties. In the present study, the effect of lactoferrin on breast cancer cell growth and the expression of Bax and Bak genes are evaluated.
Materials and Methods: MCF7 cells were cultured in a 96-well plate with 1×105 cells in each well. Different lactoferrin concentrations of 0, 50, 300, 600, and 800 μg/mL were added to each well in three replicates and the well was incubated for 24 hours. After treatment, cell survival was measured using the MTT assay. To determine the level of expression of Bax and Bak genes, the cells were treated with lactoferrin concentrations of 0, 50, and 800 μg/mL in 2 replicates for 24 hours. Then RNA extraction was performed and cDNA was synthesized immediately and the expression of the genes in the presence of beta-actin reference gene and cyber-green fluorescence color was investigated with real-time reactions.
Results: The cells viability in lactoferrin concentrations of 0, 50, 300, 500 and 800 μg/μL were 100%, 94%, 83%, 62%, and 32%, respectively. The expression level of the Bax gene at a concentration of 50 μg increased by 2.71 times and in 800 μg concentration decreased by 0.88 times. Also, the expression level of the Bak gene at concentrations of 50 and 800 μg increased by 1.23 and 1.0 fold, respectively. Statistical analysis of the data indicated that the expression levels of two genes at a concentration of 50 μg/mL of lactoferrin significantly increased (P<0.01), compared to the control. The significance level in this study was set at < 0.05.
Conclusion: In this study, lactoferrin showed a growth inhibitory effect on breast cancer cells and increased the expression of Bax and Bak genes involved in apoptosis at a concentration of 50 µg/mL.
Materials and Methods: MCF7 cells were cultured in a 96-well plate with 1×105 cells in each well. Different lactoferrin concentrations of 0, 50, 300, 600, and 800 μg/mL were added to each well in three replicates and the well was incubated for 24 hours. After treatment, cell survival was measured using the MTT assay. To determine the level of expression of Bax and Bak genes, the cells were treated with lactoferrin concentrations of 0, 50, and 800 μg/mL in 2 replicates for 24 hours. Then RNA extraction was performed and cDNA was synthesized immediately and the expression of the genes in the presence of beta-actin reference gene and cyber-green fluorescence color was investigated with real-time reactions.
Results: The cells viability in lactoferrin concentrations of 0, 50, 300, 500 and 800 μg/μL were 100%, 94%, 83%, 62%, and 32%, respectively. The expression level of the Bax gene at a concentration of 50 μg increased by 2.71 times and in 800 μg concentration decreased by 0.88 times. Also, the expression level of the Bak gene at concentrations of 50 and 800 μg increased by 1.23 and 1.0 fold, respectively. Statistical analysis of the data indicated that the expression levels of two genes at a concentration of 50 μg/mL of lactoferrin significantly increased (P<0.01), compared to the control. The significance level in this study was set at < 0.05.
Conclusion: In this study, lactoferrin showed a growth inhibitory effect on breast cancer cells and increased the expression of Bax and Bak genes involved in apoptosis at a concentration of 50 µg/mL.
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