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苦艾提取物负载聚合物纳米粒的细胞毒性及其对乳腺癌细胞的蛋白靶点研究

Cytotoxic potential of Artemisia absinthium extract loaded polymeric nanoparticles against breast cancer cells: Insight into the protein targets.

机构信息

Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi 110062, India.

Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi 110062, India.

出版信息

Int J Pharm. 2020 Aug 30;586:119583. doi: 10.1016/j.ijpharm.2020.119583. Epub 2020 Jun 27.

Abstract

Targeted drug delivery system in the form of herbal based nano-formulations is the new ray of hope for minimizing the side effects related to the anti-cancer drugs as well as conventional drug delivery system. In view of this, the present study was designed to evaluate the cytotoxic potential of A. absinthium extract loaded polymeric nanoparticles (NVA-AA) against the breast cancer cell lines (MCF-7 and MDA MB-231) and to identify the protein targets for the caused cytotoxicity. The polymeric nanoparticles (PNPs) were prepared by free radical mechanism and loaded with the whole plant extract. The cytotoxicity of these NVA-AA were evaluated on the breast cancer cell lines via different cytotoxic parameters viz. MTT assay, CFSE proliferation assay, apoptosis assay, cell cycle study. The protein targets and the interaction among them were identified by nano-LCMS/MS analysis and STRING online tool respectively, which were further validated by qPCR and BLI. The LCMS/MS analysis suggests that the caused cytotoxicity was due to the alteration of proteins involved in vesicular trafficking, apoptosis, proliferation and metastasis. Further, interactome analysis identified UBA52 in MCF-7 and TIAL1, PPP1CC in MDA MB-231 cells as the central molecule in the vesicular trafficking and apoptosis networking connection.

摘要

基于草药的纳米制剂的靶向药物输送系统是减少与抗癌药物和传统药物输送系统相关的副作用的新希望。有鉴于此,本研究旨在评估艾蒿提取物负载聚合物纳米粒(NVA-AA)对乳腺癌细胞系(MCF-7 和 MDA MB-231)的细胞毒性潜力,并鉴定引起细胞毒性的蛋白靶标。聚合物纳米粒(PNP)通过自由基机制制备并负载全植物提取物。通过不同的细胞毒性参数,即 MTT 测定、CFSE 增殖测定、凋亡测定、细胞周期研究,评估这些 NVA-AA 的细胞毒性。通过 nano-LCMS/MS 分析和 STRING 在线工具分别鉴定蛋白靶标及其相互作用,并用 qPCR 和 BLI 进一步验证。LCMS/MS 分析表明,引起的细胞毒性是由于参与囊泡转运、凋亡、增殖和转移的蛋白发生改变所致。此外,互作分析鉴定出 MCF-7 中的 UBA52 和 MDA MB-231 细胞中的 TIAL1、PPP1CC 作为囊泡转运和凋亡网络连接中的中心分子。

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