Abstract
Epidermal growth factor receptor (EGFR) overexpression is related to the increased aggressiveness, metastases, and poor prognosis in various cancers. In this study, we successfully constructed a new EGFR nanobody-based immunotoxin rE/CUS containing cucurmosin (CUS), The immunotoxin was expressed by prokaryotic system and we obtained a yield of 5 mg protein per liter expression medium. The percentage of it's binding ability totumor cell lines A549, HepG2, SW116, which highly expressed EGFR was 55.6%, 79.6% and 97.1%, respectively, but SW620 was only 4.45%. rE/CUS has the ability to bind A549, HepG2, SW116 cells specifically, and the antigen binding capability was not affected because of extra part of CUS component. The rE/CUS significantly inhibited the cell viability against EGFR over expression tumor cell lines in a dose-and time-dependent manner. Moreover, rE/CUS also induced apoptosis of HepG2 and A549 mightily. Our results demonstrate that rE/CUS is a potential therapeutic strategy for treating EGFR-positive solid tumors.
Highlights
In 2012, approximately 14 million new cancer cases were recorded and almost 8.2 million people died from cancer all around the world
We successfully constructed a new Epidermal growth factor receptor (EGFR) nanobody-based immunotoxin rE/CUS containing cucurmosin (CUS), The immunotoxin was expressed by prokaryotic system and we obtained a yield of 5 mg protein per liter expression medium
All the amplified products were transferred into agarose gel for electrophoresis (Figure 2A), the fusion gene rE/CUS was cloned into pET-32a (+) and transfected into BL21 (DE3) E.coli cells and seduced by Isopropyl β-D-1-thiogalactopyranoside (IPTG)
Summary
In 2012, approximately 14 million new cancer cases were recorded and almost 8.2 million people died from cancer all around the world. Malignant tumor has become one of the common causes of death in the US, and accounted for nearly 1 of every 4 deaths [1]. Those most fatal one are epithelial cells derived from lungs, liver, breasts, prostate, and stomach [2]. Treated with surgery, radiation, chemotherapy, and antibody-mediated receptor targeted therapy, etc., patients still suffer from unfavorable prognosis. Epidermal growth factor receptor (EGFR, ErbB-1 or HER1) over -expression can be detected on numerous epithelium-originated tumor cells, especially those generated from respiratory and digestive systems [3]. It is clear that more than two third of breast cancers express
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