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,

3,

4,

5} , Ghazala Mustafa _{3,

6,

7,

8} , Javed Iqbal _{9,

10,

11,

12} , Muhammad Ashfaq _{1,

2,

3} , Nasir Mahmood _{13,

14,

15,

16,

17}

Affiliation

Department of Biochemistry and Biotechnology
The Islamia University of Bahawalpur
Pakistan
Department of Materials Science and Engineering
College of Engineering
Department of Plant Sciences
Quaid-i-Azam University
Islamabad 45320
College of Life Sciences
Shenzhen University
Shenzhen 10590
China
School of Engineering
RMIT University
3001 Melbourne
Australia
Center of Micro-Nano Functional Materials and Devices

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细胞对生物相容性Fe ₂ C @ C纳米粒子的响应的定量蛋白质组学分析：¹⁶ O / ¹⁸ O标记和HPLC-ESI-轨道阱分析方法

在生物体内有效检测分子生物标志物，例如蛋白质，脂质，碳水化合物和病原体，是纳米医学领域的巨大挑战。在这里，我们使用细胞培养的¹⁶ O / ¹⁸ O同位素标记研究了HeLa细胞对生物相容的Fe ₂ C @ C纳米粒子（NPs）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 NPs的相对结合效率。HPLC-ESI轨道阱分析发现51种差异表达的蛋白，其中23种过表达且28种下调。这项研究表明，Fe ₂C @ C NPs改变参与胞吞作用，细胞周期调节和细胞膜突出的蛋白质的表达。此外，通过细胞色素C的Western印迹分析成功地证实了质谱（MS）结果的定量和验证。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。这项研究是对Fe ₂ C @ C NPs上低丰度蛋白的首次大规模表征，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：2017-11-24

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