牛乳铁蛋白对乳腺癌细胞系MCF7的抗癌作用及凋亡相关Bax、Bak基因表达的影响

Amir Khalafi, F. Moradian, A. Rafiei
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引用次数: 2

摘要

背景:乳铁蛋白是一种糖蛋白,具有抗菌、抗氧化、免疫调节、抗病毒和最重要的抗癌特性。本研究旨在探讨乳铁蛋白对乳腺癌细胞生长及Bax、Bak基因表达的影响。材料与方法:MCF7细胞培养于96孔板中,每孔1×105细胞。每孔添加不同浓度的乳铁蛋白,分别为0、50、300、600和800 μg/mL,分3个重复,孵育24小时。治疗后,用MTT法测定细胞存活率。为了检测Bax和Bak基因的表达水平,分别用浓度为0、50和800 μg/mL的乳铁蛋白处理细胞,分2个重复处理24小时。提取RNA,立即合成cDNA,实时反应检测基因在β -肌动蛋白内参基因和cyber-green荧光色存在下的表达情况。结果:乳铁蛋白浓度为0、50、300、500、800 μg/μL时,细胞存活率分别为100%、94%、83%、62%、32%。Bax基因在浓度为50 μg时表达量增加2.71倍,在浓度为800 μg时表达量减少0.88倍。在浓度为50 μg和800 μg时,Bak基因的表达量分别增加了1.23倍和1.0倍。数据统计分析表明,与对照组相比,乳铁蛋白浓度为50 μg/mL时,两个基因的表达量显著升高(P<0.01)。本研究的显著性水平设为< 0.05。结论:在本研究中,乳铁蛋白在浓度为50µg/mL时,对乳腺癌细胞具有生长抑制作用,并增加凋亡相关Bax和Bak基因的表达。
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Anticancer Effect of Bovine Lactoferrin on Breast Cancer Cell Line MCF7 and the Evaluation of Bax and Bak Genes Expression Involved in Apoptosis
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.
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