The tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a potent inducer of tumor cell apoptosis, but concerns of considerable liver toxicity limit its uses in human cancer therapy. Here, we show that i.v. injected Escherichia coli DH5a (E. coli DH5a) specifically replicates in solid tumors and metastases in live animals. E. coli DH5a does not enter tumor cells and suits for being the vector for soluble TRAIL (sTRAIL), which induces apoptosis by activating cell-surface death receptors. With the high 'tumor-targeting' nature, we demonstrate that intratumoral (i.t.) and intravenous injection of sTRAIL-expressing E. coli DH5a results in the tumor-targeted release of biologically active molecules, which leads to a dramatic reduction in the tumor growth rate and the prolonged survival of tumor-bearing mice. TRAIL delivery by E. coli DH5a did not cause any detectable toxicity to any organs, suggesting that E. coli DH5a-delivered sTRAIL protein therapy may provide a feasible and effective form of treatment for solid tumors.
Zhang et al.: Preparation of Soluble Protein Drug T34ATAT m -survivin (T34A), an anticancer protein drug, displayed pro-apoptotic bioactivity against various cancer cells. It was expressed in Escherichia coli as inclusion bodies. To test the bioactivity of soluble TAT msurvivin (T34A), a feasible strategy was first developed for the soluble expression and purification. Effect of zinc ion and induction temperature was investigated to improve the solubility and overall production level of TAT m -survivin (T34A). High solubility (92 %) was achieved by addition of zinc ion and temperature downshift after induction. An efficient protocol of purification was established by heat release, ammonium sulphate precipitation, anion exchange and heparin affinity chromatography. Purified TAT m -survivin (T34A) was obtained with a purity of 96 %, which inhibited the proliferation of human pancreas carcinoma cell lines SW1990. In comparison to denatured TAT m -survivin (T34A), soluble TAT m -survivin (T34A) did not exhibit higher bioactivity as expected. This study represented a novel strategy to obtain highly soluble form of TAT m -survivin (T34A) and would provide useful information for production of other soluble tat-mediated fusion proteins.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.