Alizadeh Siamak, Barzegari Abolfazl, Esmaeili Abolghasem, Omidi Yadollah
Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran.
Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran.
Bioimpacts. 2020;10(3):187-193. doi: 10.34172/bi.2020.23. Epub 2019 Nov 2.
Colorectal cancer (CRC) is one of the main health burden worldwide, which can cause major economic and physiological problems along with relatively high rate of mortality. It is important to develop new methods for the localized delivery of recombinant protein therapeutics, in large part due to the failure of conventional therapies in most cases. Since Nissle 1917 (EcN) does not produce any virulence factors, here we used these bacteria with the light-activated promoter system to deliver therapeutic agents in the desired location and time. In this study, alpha hemolysin (SAH), after codon usage optimization, was cloned into blue light activating vector (pDawn) and transferred to EcN strain. Then, the functionality and cytotoxicity of secreted alpha hemolysin was evaluated in the SW480 colon cancer cell line by using different experiments, including blood agar test, flow cytometry analysis, and DAPI staining. Our findings revealed that EcN can produce functional SAH under the blue light irradiation against SW480 cancer cells. Moreover, cytotoxicity assays confirmed the dose- and time-dependent toxicity of this payload (SAH) against SW480 cancer cells. Based on our results, EcN is proposed as an appropriate light-activated vehicle for delivery of anticancer agents to the target cancer cells/tissues.
结直肠癌（CRC）是全球主要的健康负担之一，它会引发重大的经济和生理问题，死亡率也相对较高。开发重组蛋白治疗药物局部递送的新方法很重要，这在很大程度上是因为传统疗法在大多数情况下效果不佳。由于Nissle 1917（EcN）不产生任何毒力因子，我们在此使用这些细菌与光激活启动子系统，在期望的位置和时间递送治疗剂。在本研究中，经过密码子优化后的α溶血素（SAH）被克隆到蓝光激活载体（pDawn）中，并转移到EcN菌株。然后，通过不同实验，包括血琼脂试验、流式细胞术分析和DAPI染色，在SW480结肠癌细胞系中评估分泌的α溶血素的功能和细胞毒性。我们的研究结果表明，EcN在蓝光照射下可产生针对SW480癌细胞的功能性SAH。此外，细胞毒性试验证实了这种负载（SAH）对SW480癌细胞具有剂量和时间依赖性毒性。基于我们的结果，EcN被认为是一种将抗癌药物递送至靶癌细胞/组织的合适的光激活载体。
