Neuropilin-1 Binding Peptide as Fusion to Diphtheria Toxin Induces Apoptosis in Non-small Cell Lung Cancer Cell Line

IF 2.6 4区医学 Q2 PHARMACOLOGY & PHARMACY

Current pharmaceutical design Pub Date : 2024-04-05 DOI:10.2174/0113816128292382240325074032

Sara Eghtedari, Mahdi Behdani, Fatemeh Kazemi-Lomedasht

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引用次数: 0

Abstract

Background:: Targeted cancer therapy can be considered as a new strategy to overcome the side effects of current cancer treatments. Neuropilin-1 (NRP-1) is a transmembrane glycoprotein that is expressed in endothelial cells and tumor vessels to stimulate angiogenesis progression. Targeted diphtheria toxin (DT)- based therapeutics are promising tools for cancer treatment. This study aimed to construct a novel NRP-1 binding peptide (as three repeats) (CRGDK) as a fusion to truncated DT (DTA) (DTA-triCRGDK) for targeted delivery of DT into NRP-1 expressing cells. Methods:: The concept of DTA-triCRGDK was designed, synthesized and cloned into the bacterial host. Expression of DTA-triCRGDK was induced by Isopropyl ß-D-1-thiogalactopyranoside (IPTG) and purification was performed using Ni-NTA chromatography. Biological activity of DTA-triCRGDK was evaluated using MTT, apoptosis, and wound healing assays. In addition, expression levels of apoptotic Bax, Bcl2, and Casp3 genes were determined by Real-time PCR Results:: Cytotoxicity analysis showed the IC50 values of DTA-triCRGDK for A549 and MRC5 were 0.43 nM and 4.12 nM after 24h, respectively. Bcl2 expression levels decreased 0.4 and 0.72 fold in A549 and MRC5, respectively. However, Bax and Casp3 expression level increased by 6.75 and 8.19 in A549 and 2.51 and 3.6 in MRC5 cells. Conclusion:: Taken together, DTA-triCRGDK is a promising tool for targeted therapy of NRP-1 overexpressing cancer cells.

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与白喉毒素融合的神经肽-1 结合肽诱导非小细胞肺癌细胞株凋亡

背景癌症靶向治疗可被视为克服当前癌症治疗副作用的一种新策略。神经蛋白-1（NRP-1）是一种跨膜糖蛋白，在内皮细胞和肿瘤血管中表达，刺激血管生成。基于白喉毒素（DT）的靶向疗法是治疗癌症的有效手段。本研究旨在构建一种新型 NRP-1 结合肽（三个重复序列）（CRGDK），作为截短 DT（DTA）（DTA-triCRGDK）的融合物，将 DT 靶向递送至 NRP-1 表达细胞。方法设计、合成 DTA-triCRGDK 的概念，并将其克隆到细菌宿主中。用异丙基ß-D-1-硫代半乳糖苷（IPTG）诱导DTA-triCRGDK的表达，并用Ni-NTA色谱法进行纯化。使用 MTT、细胞凋亡和伤口愈合试验评估了 DTA-triCRGDK 的生物活性。此外，还通过实时 PCR 检测了凋亡 Bax、Bcl2 和 Casp3 基因的表达水平：细胞毒性分析表明，24 小时后，DTA-triCRGDK 对 A549 和 MRC5 的 IC50 值分别为 0.43 nM 和 4.12 nM。A549 和 MRC5 中的 Bcl2 表达水平分别下降了 0.4 和 0.72 倍。然而，Bax 和 Casp3 的表达水平在 A549 细胞中分别增加了 6.75 倍和 8.19 倍，在 MRC5 细胞中分别增加了 2.51 倍和 3.6 倍。结论综上所述，DTA-triCRGDK 是一种很有前景的靶向治疗 NRP-1 过表达癌细胞的工具。

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来源期刊

Current pharmaceutical design 医学-药学

CiteScore

6.30

自引率

0.00%

发文量

302

审稿时长

2 months

期刊介绍： Current Pharmaceutical Design publishes timely in-depth reviews and research articles from leading pharmaceutical researchers in the field, covering all aspects of current research in rational drug design. Each issue is devoted to a single major therapeutic area guest edited by an acknowledged authority in the field. Each thematic issue of Current Pharmaceutical Design covers all subject areas of major importance to modern drug design including: medicinal chemistry, pharmacology, drug targets and disease mechanism.