Quantitative proteomic analysis of HeLa cells in response to biocompatible Fe2C@C nanoparticles: 16O/18O-labelling & HPLC-ESI-orbit-trap profiling approach†,Toxicology Research

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Quantitative proteomic analysis of HeLa cells in response to biocompatible Fe2C@C nanoparticles: 16O/18O-labelling & HPLC-ESI-orbit-trap profiling approach†
Toxicology Research ( IF 2.2 ) Pub Date : 2017-11-08 00:00:00 , DOI: 10.1039/c7tx00248c
Murtaza Hasan _{1,

2} , Ghazala Mustafa ₃ , Javed Iqbal ₄ , Muhammad Ashfaq ₁ , Nasir Mahmood _{5,

6}

Affiliation

The effective detection of molecular biomarkers, such as proteins, lipids, carbohydrates, and pathogens, in a living body is a huge challenge in the field of nanomedicine. Here, we have investigated the comparative quantitative proteomics analysis of the molecular response of HeLa cells to biocompatible Fe₂C@C nanoparticles (NPs) using ¹⁶O/¹⁸O isotopic labelling of the cell culture. The relative binding efficiency of proteins to Fe₂C@C NPs was calculated. HPLC-ESI-orbit-trap analysis found 51 differentially expressed proteins, out of which 23 were over-expressed and 28 down-regulated. This study showed that Fe₂C@C NPs alter the expression of the proteins involved in endocytosis, cell-cycle regulation, and cell membrane protrusion. Further, the quantification and validation of the mass spectrometry (MS) results was successfully confirmed by western blot analysis of cytochrome C. The change in the expression of proteins can be useful for early stage disease diagnoses and the development of tailored therapeutic strategies. This study is the first large-scale characterization of low abundance proteins on Fe₂C@C NPs, providing the biochemical basis for the assessment of the suitability of magnetic NPs as biomedical markers and emerging functional probes.

中文翻译：

HeLa 细胞响应生物相容性 Fe2C@C 纳米粒子的定量蛋白质组分析：16O/18O 标记和 HPLC-ESI-orbit-trap 分析方法†

有效检测活体中的蛋白质、脂质、碳水化合物和病原体等分子生物标志物是纳米医学领域的巨大挑战。在这里，我们使用细胞培养物的¹⁶ O/ ¹⁸ O 同位素标记，研究了 HeLa 细胞对生物相容性 Fe ₂ C@C 纳米颗粒 (NP) 的分子响应的比较定量蛋白质组学分析。计算了蛋白质与 Fe ₂ C@C NP 的相对结合效率。 HPLC-ESI-orbit-trap 分析发现 51 个差异表达蛋白，其中 23 个过表达，28 个下调。这项研究表明，Fe ₂ C@C NPs 改变了参与内吞作用、细胞周期调节和细胞膜突出的蛋白质的表达。此外，细胞色素 C 的蛋白质印迹分析成功证实了质谱 (MS) 结果的量化和验证。蛋白质表达的变化可用于早期疾病诊断和定制治疗策略的开发。这项研究是Fe ₂ C@C NPs上低丰度蛋白质的首次大规模表征，为评估磁性NPs作为生物医学标记和新兴功能探针的适用性提供了生化基础。

更新日期：2017-11-08

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