Ghasemi-Dehkordi Payam, Doosti Abbas, Jami Mohammad-Saeid
Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran.
Saudi J Biol Sci. 2020 Sep;27(9):2308-2317. doi: 10.1016/j.sjbs.2020.04.007. Epub 2020 Apr 14.
Azurin protein of 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 and human 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 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 and genes in the MCF-7 cell were confirmed by RT-PCR, IF and western blotting. The apoptosis assay was showed a statistically significant ( < 0.05) difference after transfection. The expression of , , and genes in transformed cells compare with non-transformed and transformed MCF-7 by pBudCE4.1 were increased statistically significant ( < 0.05) increases. Although, the increase of and expressions in transformed cells were not statistically significant ( > 0.05). Co-expression of and genes could induce apoptosis and necrosis in human MCF-7 breast cancer cells by up-regulation of , and 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.
天青蛋白是一种抗乳腺癌的抗肿瘤药物,而乳腺珠蛋白-A(MAM-A)蛋白是MCF-7细胞表面用于诱导细胞免疫的特异性抗原。本研究的目的是探讨[此处原文缺失基因名称]和人[此处原文缺失基因名称]基因的同时表达对MCF-7乳腺癌细胞系中凋亡相关基因和细胞周期基因mRNA表达水平的影响。使用脂质体转染试剂将重组质粒或空质粒分别转入MCF-7细胞。通过流式细胞术检测细胞死亡和凋亡情况。采用免疫荧光分析、逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法评估[此处原文缺失基因名称]基因的表达。最后,通过定量聚合酶链反应(qPCR)方法检测转化和未转化的MCF-7细胞中凋亡相关基因和细胞周期基因的表达。通过RT-PCR、免疫荧光和蛋白质免疫印迹法证实了[此处原文缺失基因名称]和[此处原文缺失基因名称]基因在MCF-7细胞中的成功表达。转染后凋亡检测显示出统计学上的显著差异(P<0.05)。与未转化的MCF-7细胞以及用pBudCE4.1载体转化的MCF-7细胞相比,转化细胞中[此处原文缺失基因名称]、[此处原文缺失基因名称]和[此处原文缺失基因名称]基因的表达有统计学意义的显著增加(P<0.05)。然而,转化细胞中[此处原文缺失基因名称]和[此处原文缺失基因名称]表达的增加无统计学意义(P>0.05)。[此处原文缺失基因名称]和[此处原文缺失基因名称]基因的共表达可通过上调[此处原文缺失基因名称]、[此处原文缺失基因名称]和[此处原文缺失基因名称]基因诱导人MCF-乳腺癌细胞凋亡和坏死。在未来的研究中,必须在动物模型中更好地评估pBudCE4.1-天青蛋白-MAM-A重组载体的免疫刺激作用,并对治疗方法进行评估。