Intestinal probiotics E. coli Nissle 1917 as a targeted vehicle for delivery of p53 and Tum-5 to solid tumors for cancer therapy
Intestinal probiotics E. coli Nissle 1917 as a targeted vehicle for delivery of p53 and Tum-5 to solid tumors for cancer therapy
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DOI：
10.1186/s13036-019-0189-9
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发表时间：
2019-06-28
影响因子：
5.6
通讯作者：
Xia, Liqiu
中科院分区：
文献类型：
--
作者：
He, Lian;Yang, Huijun;Xia, Liqiu
传统的肿瘤治疗方法，如手术治疗、放疗、化疗等，往往不能完全清除肿瘤区域缺氧微环境中的肿瘤细胞，与这些传统的肿瘤治疗方法相比，利用靶向递送载体将抗癌基因或抗癌药物递送到肿瘤缺氧区域是近年来发展迅速的最具临床应用前景的肿瘤治疗方法。在这项研究中，大肠杆菌Nissle 1917（EcN），一种肠道益生菌，被用作靶向转运载体，将p53和Tum-5蛋白递送到肿瘤缺氧区域。利用荧光素酶LuxCDABE操纵子研究了EcN的肿瘤靶向性，结果表明EcN可特异性聚集于荷SMMC-7721裸鼠的实体瘤区域。首先构建Tum 5-p53双功能蛋白，然后通过使用靶向转运蛋白EcN将其递送到实体瘤区域用于癌症治疗。同时对EcN（Tum-5）、EcN（p53）和EcN（Tum 5-p53）三种工程菌的抗肿瘤作用和安全性进行了研究。计算肿瘤体积和瘤重表明，3种工程菌均能抑制人肝癌SMMC-7721细胞的生长，且表达Tum 5-p53融合蛋白的EcN（Tum 5-p53）的抗肿瘤效果明显优于单独的EcN（Tum-5）和EcN（p53）。免疫荧光检测显示，工程菌处理组肿瘤区域核增殖相关蛋白Ki-67的表达受到抑制，而caspase-3的表达上调。Ki-67和caspase-3的表达趋势与3种工程菌不同的抗肿瘤效果相一致。EcN对小鼠无明显毒副作用。本研究不仅为肿瘤的靶向治疗奠定了基础，而且对抗肿瘤药物和抗肿瘤蛋白的开发具有重要意义。
Traditional cancer therapies, such as surgery treatment, radiotherapy, and chemotherapy, often fail to completely eliminate tumor cells in an anaerobic microenvironment of tumor regions.In contrast to these traditional cancer therapies, the use of targeted delivery vectors to deliver anticancer genes or antitumor drugs to hypoxic areas in tumors is the most clinically promising cancer treatment with rapid development in recent years. In this study, E.coli Nissle 1917 (EcN), an intestinal probiotic, was utilized as a targeted transport vector to deliver p53 and Tum-5 protein to tumor hypoxic regions. The tumor-targeting characteristics of EcN were investigated using luciferase LuxCDABE operon, and the results demonstrated that EcN could specifically accumulate in the solid tumor areas of SMMC-7721 tumor-bearing BALB/c nude mice. The Tum 5-p53 bifunctional proteins were initially constructed and then delivered to solid tumor regions by using the targeted transporter EcN for cancer therapy. The antitumor effect and safety of three engineered bacteria, namely, EcN (Tum-5), EcN (p53), and EcN (Tum 5-p53), were also examined. The calculated tumor volume and tumor weight indicated that these three engineered bacteria could inhibit the growth of human hepatoma SMMC-7721 cells, and the antitumor effect of EcN (Tum 5-p53) expressing the Tum 5-p53 fusion protein was significantly better than those of EcN (Tum-5) and EcN (p53) alone. Immunofluorescence demonstrated that the expression of Ki-67, a nuclear proliferation-related protein, was inhibited in the tumor areas of the groups treated with the engineered bacteria, whereas the expression of caspase-3 was upregulated. The expression trends of Ki-67 and caspase-3 were consistent with the different antitumor efficacies of these three engineered bacteria. EcN did not elicit obvious side effects on mice. This research not only provids a foundation for tumor-targeted therapy but also contributes greatly to the development of antitumor agents and anticancer proteins.