大肠杆菌MF-107中L-天冬氨酸酶对MCF7细胞株抗肿瘤作用的生化和生物学评价

Q2 Biochemistry, Genetics and Molecular Biology
Masoumeh Shahnazari, R. Bigdeli, A. Dashbolaghi, Reza Ahangari Cohan, Alireza Shoari, H. Hosseini, Davoud Nouri Inanlou, V. Asgary
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引用次数: 1

摘要

背景:微生物L-ASNase是应用最广泛的抗癌药物之一。在此,我们评估了从革兰氏阴性菌菌株大肠杆菌MF-107分离的L-ASNase的生化和生物学特性。方法:以芦笋为试材,从中分离出多株细菌。进一步筛选这些菌株的L-ASNase活性。选择了一种有前景的细菌分离物用于L-ASNase的生产和随后的纯化。测定了纯化的L-ASNase的分子量。应用MTT法评估纯化的酶的细胞毒性作用。此外,为了测定胱天蛋白酶活性和纯化酶对细胞内凋亡的影响,我们进行了实时PCR方法。结果:该酶的分子量约为37kDa。在7.5至8的pH范围内,该酶具有相当大的稳定性。在35°C下,记录纯化的L-ASNase的最佳活性。酶对处理细胞的细胞毒性作用是剂量依赖性的,IC50值为5.7IU/ml。L-ASNase治疗后Bax基因表达显著增加5.75倍(p<0.001)。另一方面,与对照组相比,抗凋亡Bcl-2基因的表达增加了2.63倍(p<0.05)。检测到胱天蛋白酶-3和p53的mRNA水平显著上调(分别为5.93和1.85倍)。与未处理的细胞相比,我们没有发现处理的细胞的胱天蛋白酶-8活性有任何变化。结论:在本研究中,从微生物来源分离的L-ASNase的细胞毒性作用显著抑制了癌症细胞的增殖。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Biochemical and Biological Evaluation of an L-Asparaginase from Isolated Escherichia coli MF-107 as an Anti-Tumor Enzyme on MCF7 Cell Line
Background: One of the most widely used anticancer agents is microbial L-ASNase. Herein, we assessed the biochemical and biological properties of an isolated L-ASNase from a Gram-negative bacteria strain, Escherichia coli MF-107. Methods: Using garden asparagus, we obtained several bacterial isolates. These strains were further screened for L-ASNase activity. A promising bacterial isolate was selected for L-ASNase production and subsequent purification. The molecular weight of purified L-ASNase was determined. The MTT assay was applied to assess the cytotoxic effect of the purified enzyme. Also, for caspase activity determination and the apoptotic effect of purified enzyme on in cells, we conducted a real-time PCR method. Results: The molecular weight of the enzyme was approximately 37 kDa. In the pH range of 7.5 to 8, the enzyme had considerable stability. At 35 °C, the purified L-ASNase optimum activity was recorded. The cytotoxic effect of the enzyme on treated cells was dose-dependent with an IC50 value of 5.7 IU/ml. The Bax gene expression considerably raised by 5.75-fold (p < 0.001) upon L-ASNase treatment. On the other hand, the anti-apoptotic Bcl-2 gene expression showed a 2.63-fold increase compared to the control (p < 0.05). It was detected that the mRNA levels of caspase-3 and p53 were considerably upregulated (5.93 and 1.85-fold, respectively). We did not find any alternation in the caspase-8 activity of the treated cells compared to untreated cells. Conclusion: In this research, the proliferation of the breast cancer cells remarkably inhibited via the cytotoxic effect of isolated L-ASNase from microbial sources.
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来源期刊
Iranian Biomedical Journal
Iranian Biomedical Journal Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.20
自引率
0.00%
发文量
42
审稿时长
8 weeks
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