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Plasma membrane proteomic analysis of human osteosarcoma and osteoblastic cells: revealing NDRG1 as a marker for osteosarcoma

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Tumor Biology

Abstract

Osteosarcoma (OS) is the most common primary malignant bone tumor in children and adolescents. To identify new biomarkers for early diagnosis of OS and novel therapeutic candidates, we carried out a plasma membrane proteomic study based on two-dimensional electrophoresis (2DE). The OS cell line MG-63 and the human osteoblastic cell line hFOB1.19 were adopted as the comparison model. We extracted plasma membrane by aqueous two-phase partition extraction. The proteins were separated through 2DE. We analyzed the differentially expressed proteins by Imagemaster software and then identified them by liquid chromatography–tandem mass spectrometry, and the location and function of differential proteins were searched through the Gene Ontology database. In total, 220 protein spots were separated by 2DE. Seven proteins with more than 2.0-folds of difference were successfully identified from 13 gel spots, with 6 up-regulated and 1 down-regulated. Gene Ontology analysis of the differentially expressed proteins indicated that these proteins were involved in seven kinds of functions including binding, structural, cell motility, receptor activity, electron carrier activity, NADH dehydrogenase (ubiquinone) activity, and transcription repressor activity. The up-regulation of NDRG1 was verified in osteosarcoma through Western blotting and by immunohistochemistry in paraffin-embedded tissues. The plasma membrane proteins identified in this study may provide new insights into osteosarcoma cancer biology and potential diagnostic and therapeutic biomarkers.

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Abbreviations

OS:

Osteosarcoma

2DE:

Two-dimensional electrophoresis

LC–MS:

Liquid chromatography–mass spectrometry

MS/MS:

Tandem MS

HCT:

High capacity trap

HE:

Hematoxylin and eosin

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Acknowledgments

We thank the grants from the National 973 Project of China (2011CB910700), Shanghai Natural Science Foundation (09ZR1426300), and Shanghai Hospital Clinical Research Platform for Clinical Research Source (No. SHDC12007203). We thank Kirill Gorshkov for carefully editing the whole manuscript.

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Authors and Affiliations

  1. Musculoskeletal Oncology Center, Shanghai 10th People’s Hospital, Tongji University School of Medicine, Shanghai, 200072, China

    Yingqi Hua, Mengxiong Sun, Longpo Zheng & Zhengdong Cai

  2. Shanghai Public Health Clinical Center affiliated to Fudan University, Shanghai, 201508, China

    Xiaofang Jia, Lin Yin & Lijun Zhang

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  1. Yingqi Hua
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  2. Xiaofang Jia
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  3. Mengxiong Sun
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  5. Lin Yin
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  6. Lijun Zhang
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  7. Zhengdong Cai
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Corresponding authors

Correspondence to Lijun Zhang or Zhengdong Cai.

Additional information

Yingqi Hua, Xiaofang Jia, Mengxiong Sun, and Longpo Zheng contributed equally to this work.

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Figure S1

The SDS-PAGE gels of 50 ug MG and Normal PM proteins to make sure equal protein load for western blotting analysis. (JPEG 15 kb)

High resolution image (TIFF 887 kb)

Table S1

Table S1 Lists of non-redundant peptides identified in each protein spot. (XLS 24 kb)

Table S2

Information of proteins involved in protein-protein interaction network. (XLS 113 kb)

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Hua, Y., Jia, X., Sun, M. et al. Plasma membrane proteomic analysis of human osteosarcoma and osteoblastic cells: revealing NDRG1 as a marker for osteosarcoma. Tumor Biol. 32, 1013–1021 (2011). https://doi.org/10.1007/s13277-011-0203-4

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  • DOI: https://doi.org/10.1007/s13277-011-0203-4

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