Alirahimi E, Ashkiyan A, Kazemi-Lomedasht F, Azadmanesh K, Hosseininejad-Chafi M, Habibi-Anbouhi M, Moazami R, Behdani M
Biotechnology Research Center, Venom & Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, Iran.
Virology Department, Pasteur Institute of Iran, Tehran, Iran.
Cancer Gene Ther. 2017 Jan;24(1):33-37. doi: 10.1038/cgt.2016.76. Epub 2016 Dec 16.
Angiogenesis is among the most important mechanisms that helps cancer cells to survive, grow and undergo metastasis. Therefore, inhibiting angiogenesis will suppress tumor growth. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) are believed to be important players of angiogenesis. The goal of this study was to evaluate the success of a novel nanobody against VEGFR2 in tethering its target inside the endoplasmic reticulum and preventing its transport to the cell membrane. Nanobody sequence was cloned in a mammalian vector in fusion with green fluorescent protein and a KDEL retention signal. After transfection of 293KDR cells with this expression vector, surface localization of VEGFR2 was monitored by flow cytometry. This study demonstrates that our intrananobody is effective in targeting VEGFR2 receptor, and therefore, it is a powerful tool to downregulate a surface-exposed target protein, and in this capacity, it has potential to be used as a therapeutic protein to inhibit growth of tumors.
血管生成是帮助癌细胞存活、生长和发生转移的最重要机制之一。因此，抑制血管生成将抑制肿瘤生长。血管内皮生长因子（VEGF）及其受体（VEGFR）被认为是血管生成的重要参与者。本研究的目的是评估一种针对VEGFR2的新型纳米抗体在将其靶标拴在内质网内并阻止其转运到细胞膜方面的成效。纳米抗体序列克隆到与绿色荧光蛋白和KDEL滞留信号融合的哺乳动物载体中。用该表达载体转染293KDR细胞后，通过流式细胞术监测VEGFR2的表面定位。本研究表明，我们的细胞内纳米抗体在靶向VEGFR2受体方面是有效的，因此，它是下调表面暴露靶蛋白的有力工具，并且以此能力，它有潜力用作抑制肿瘤生长的治疗性蛋白。
