Top

Published in:

01-10-2011 | Research Article

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

Authors: Yingqi Hua, Xiaofang Jia, Mengxiong Sun, Longpo Zheng, Lin Yin, Lijun Zhang, Zhengdong Cai

Published in: Tumor Biology | Issue 5/2011

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.

Available only for authorised users

Mankin HJ, Hornicek FJ, Rosenberg AE, Harmon DC, Gebhardt MC. Survival data for 648 patients with osteosarcoma treated at one institution. Clin Orthop Relat Res. 2004;(429):286–91.

Leth-Larsen R, Lund RR, Ditzel HJ. Plasma membrane proteomics and its application in clinical cancer biomarker discovery. Mol Cell Proteomics. 2010;9:1369–82.PubMedCrossRef

Sprenger RR, Jensen ON. Proteomics and the dynamic plasma membrane: quo vadis? Proteomics. 2010;10:3997–4011.PubMedCrossRef

Bhattacharyya S, Byrum S, Siegel ER, Suva LJ. Proteomic analysis of bone cancer: a review of current and future developments. Expert Rev Proteomics. 2007;4:371–8.PubMedCrossRef

Byrum S, Montgomery CO, Nicholas RW, Suva LJ. The promise of bone cancer proteomics. Ann N Y Acad Sci. 2010;1192:222–9.PubMedCrossRef

Folio C, Mora MI, Zalacain M, Corrales FJ, Segura V, Sierrasesumaga L, et al. Proteomic analysis of chemonaive pediatric osteosarcomas and corresponding normal bone reveals multiple altered molecular targets. J Proteome Res. 2009;8:3882–8.PubMedCrossRef

Guo QC, Shen JN, Jin S, Wang J, Huang G, Zhang LJ, et al. Comparative proteomic analysis of human osteosarcoma and sv40-immortalized normal osteoblastic cell lines. Acta Pharmacol Sin. 2007;28:850–8.PubMedCrossRef

Zhang Z, Zhang L, Hua Y, Jia X, Li J, Hu S, et al. Comparative proteomic analysis of plasma membrane proteins between human osteosarcoma and normal osteoblastic cell lines. BMC Cancer. 2010;10:206.PubMedCrossRef

Cao R, Li X, Liu Z, Peng X, Hu W, Wang X, et al. Integration of a two-phase partition method into proteomics research on rat liver plasma membrane proteins. J Proteome Res. 2006;5:634–42.PubMedCrossRef

10.

Zhang L, Jia X, Zhang X, Sun J, Peng X, Qi T, et al. Proteomic analysis of PBMCs: characterization of potential HIV-associated proteins. Proteome Sci. 2010;8:12.PubMedCrossRef

11.

Zhang L, Jia X, Zhang X, Cao J, Yang P, Qiu C, et al. Alpha-1 antitrypsin variants in plasma from HIV-infected patients revealed by proteomic and glycoproteomic analysis. Electrophoresis. 2010;31:3437–45.PubMedCrossRef

12.

Zhang L, Xie J, Wang X, Liu X, Tang X, Cao R, et al. Proteomic analysis of mouse liver plasma membrane: use of differential extraction to enrich hydrophobic membrane proteins. Proteomics. 2005;5:4510–24.PubMedCrossRef

13.

de Alava E. Molecular pathology in sarcomas. Clin Transl Oncol. 2007;9:130–44.PubMedCrossRef

14.

Dass CR, Ek ET, Choong PF. PEDF as an emerging therapeutic candidate for osteosarcoma. Curr Cancer Drug Targets. 2008;8:683–90.PubMedCrossRef

15.

Nikitovic D, Berdiaki K, Chalkiadaki G, Karamanos N, Tzanakakis G. The role of SLRP-proteoglycans in osteosarcoma pathogenesis. Connect Tissue Res. 2008;49:235–8.PubMedCrossRef

16.

Zhang L, Peng X, Zhang Z, Feng Y, Jia X, Shi Y, et al. Subcellular proteome analysis unraveled annexin A2 related to immune liver fibrosis. J Cell Biochem. 2010;110:219–28.PubMed

17.

Zhang L, Jia X, Feng Y, Peng X, Zhang Z, Zhou W, et al. Plasma membrane proteome analysis of the early effect of alcohol on liver: implications for alcoholic liver disease. Acta Biochim Biophys Sin (Shanghai). 2011;43:19–29.CrossRef

18.

Lubke T, Lobel P, Sleat DE. Proteomics of the lysosome. Biochim Biophys Acta. 2009;1793:625–35.PubMedCrossRef

19.

Jung EU, Yoon JH, Lee YJ, Lee JH, Kim BH, Yu SJ, et al. Hypoxia and retinoic acid-inducible NDRG1 expression is responsible for doxorubicin and retinoic acid resistance in hepatocellular carcinoma cells. Cancer Lett. 2010;298:9–15.PubMedCrossRef

20.

Sibold S, Roh V, Keogh A, Studer P, Tiffon C, Angst E, et al. Hypoxia increases cytoplasmic expression of NDRG1, but is insufficient for its membrane localization in human hepatocellular carcinoma. FEBS Lett. 2007;581:989–94.PubMedCrossRef

21.

Angst E, Sibold S, Tiffon C, Weimann R, Gloor B, Candinas D, et al. Cellular differentiation determines the expression of the hypoxia-inducible protein NDRG1 in pancreatic cancer. Br J Cancer. 2006;95:307–13.PubMedCrossRef

22.

Cangul H. Hypoxia upregulates the expression of the NDRG1 gene leading to its overexpression in various human cancers. BMC Genet. 2004;5:27.PubMedCrossRef

23.

Gerhard R, Nonogaki S, Fregnani JH, Soares FA, Nagai MA. Ndrg1 protein overexpression in malignant thyroid neoplasms. Clinics (Sao Paulo). 2010;65:757–62.

24.

Matsugaki T, Zenmyo M, Hiraoka K, Fukushima N, Shoda T, Komiya S, et al. N-myc downstream-regulated gene 1/Cap43 expression promotes cell differentiation of human osteosarcoma cells. Oncol Rep. 2010;24:721–5.PubMed

25.

Wu M, Bai X, Xu G, Wei J, Zhu T, Zhang Y, et al. Proteome analysis of human androgen-independent prostate cancer cell lines: variable metastatic potentials correlated with vimentin expression. Proteomics. 2007;7:1973–83.PubMedCrossRef

26.

Hayashi E, Kuramitsu Y, Okada F, Fujimoto M, Zhang X, Kobayashi M, et al. Proteomic profiling for cancer progression: differential display analysis for the expression of intracellular proteins between regressive and progressive cancer cell lines. Proteomics. 2005;5:1024–32.PubMedCrossRef

27.

He X, Cao X. Identification of alternatively spliced GRIM-19 mRNA in kidney cancer tissues. J Hum Genet. 2010;55:507–11.PubMedCrossRef

28.

Li Y, Liang Q, Wen YQ, Chen LL, Wang LT, Liu YL, et al. Comparative proteomics analysis of human osteosarcomas and benign tumor of bone. Cancer Genet Cytogenet. 2010;198:97–106.PubMedCrossRef

29.

Shimizu T, Chiqira M, Nagase M, Watanabe H, Udagawa E. HLA phenotypes in patients who have osteosarcoma. J Bone Jt Surg Am. 1990;72:68–70.

Title: Plasma membrane proteomic analysis of human osteosarcoma and osteoblastic cells: revealing NDRG1 as a marker for osteosarcoma
Authors: Yingqi Hua
Xiaofang Jia
Mengxiong Sun
Longpo Zheng
Lin Yin
Lijun Zhang
Zhengdong Cai
Publication date: 01-10-2011
Publisher: Springer Netherlands
Published in: Tumor Biology / Issue 5/2011
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI: https://doi.org/10.1007/s13277-011-0203-4

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 5/2011

Hypoxia-related protein expression and its clinicopathologic implication in carcinoma of unknown primary

Erratum to: Promoter hypermethylation of RASSF1A, MGMT, and HIC-1 genes in benign and malignant colorectal tumors

Involvement of ERK1/2/NF-κB signal transduction pathway in TF/FVIIa/PAR2-induced proliferation and migration of colon cancer cell SW620

Intracellular clusterin negatively regulates ovarian chemoresistance: compromised expression sensitizes ovarian cancer cells to paclitaxel

Quantitative expression analysis of apoptotic/antiapoptotic genes and association with immunolocalization of BAX and BCL-2 in peripheral and central giant cell lesions of the jaws

The nucleosome binding protein NSBP1 is highly expressed in human bladder cancer and promotes the proliferation and invasion of bladder cancer cells

Keynote webinar | Spotlight on antibody–drug conjugates in cancer