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Updated: Jul 11, 2026
Flow-sorting and Exome Sequencing of the Reed-Sternberg Cells of Classical Hodgkin Lymphoma
Published on: June 10, 2017
Comprehensive identification of proteins in Hodgkin lymphoma-derived Reed-Sternberg cells by LC-MS/MS
Jeremy C Wallentine1, Ki Kwon Kim, Charles E Seiler
1Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.
Insights
This study identified 1945 proteins in Hodgkin lymphoma cells and released proteins, revealing potential biomarkers for disease understanding and pathogenesis. Proteomics analysis offers a high-throughput approach for cancer research.
Area of Science:
- Proteomics
- Cancer Biology
- Biochemistry
Background:
- Hodgkin lymphoma is a malignancy characterized by Reed-Sternberg cells.
- Understanding the proteome of these cells is crucial for biomarker discovery and pathogenesis insights.
- Previous proteomic analyses of Hodgkin lymphoma have been limited in scope.
Purpose of the Study:
- To comprehensively identify and characterize the proteome of Hodgkin lymphoma-derived Reed-Sternberg cells.
- To analyze proteins expressed in different cellular compartments (cytosolic, membrane, nuclear) and those released by the cells.
- To discover potential protein biomarkers for Hodgkin lymphoma.
Main Methods:
- Subcellular fractionation of L428 and KMH2 Hodgkin lymphoma cell lines.
- Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for proteomic analysis.
- Bioinformatics analysis using TurboSEQUEST against the UniProt database.
- Validation of selected proteins by Western blot, immunofluorescence microscopy, and immunohistochemistry.
Main Results:
- Identification of a total of 1945 proteins, including 785 cytosolic, 305 membrane, 441 nuclear, and 414 released proteins.
- Proteins identified span diverse functional groups, reflecting the complexity of the Reed-Sternberg cell proteome.
- Validated expression of selected proteins (e.g., B-RAF, Erb-B3, Axin1, Tenascin-X, Mucin-2, PIM1) in Hodgkin lymphoma cells and tissues.
Conclusions:
- This study provides the first comprehensive inventory of proteins expressed by Reed-Sternberg cells in Hodgkin lymphoma.
- The findings highlight the utility of integrated proteomic approaches (subfractionation, LC-MS/MS, bioinformatics) for biomarker discovery.
- Identified proteins offer potential as diagnostic or prognostic biomarkers and warrant further investigation into their role in Hodgkin lymphoma pathogenesis.
Abstract:
Mass spectrometry-based proteomics in conjunction with liquid chromatography and bioinformatics analysis provides a highly sensitive and high-throughput approach for the identification of proteins. Hodgkin lymphoma is a form of malignant lymphoma characterized by the proliferation of Reed-Sternberg cells and background reactive lymphocytes. Comprehensive analysis of proteins expressed and released by Reed-Sternberg cells would assist in the discovery of potential biomarkers and improve our understanding of its pathogenesis. The subcellular proteome of the three cellular compartments from L428 and KMH2 Hodgkin lymphoma-derived cell lines were fractionated, and analyzed by reverse-phase liquid chromatography coupled with electrospray ionization tandem mass spectrometry. Additionally, proteins released by Hodgkin lymphoma-derived L428 cells were extracted from serum-free culture media and analyzed. Peptide spectra were analyzed using TurboSEQUEST against the UniProt protein database (5.26.05; 188 712 entries). A subset of the identified proteins was validated by Western blot analysis, immunofluorescence microscopy and immunohistochemistry. A total of 1945 proteins were identified with 785 from the cytosolic fraction, 305 from the membrane fraction, 441 from the nuclear fraction and 414 released proteins using a minimum of two peptide identifications per protein and an error rate of <5.0%. Identification of proteins from diverse functional groups reflected the functional complexity of the Reed-Sternberg proteome. Proteins with previously reported oncogenic function in other cancers and from signaling pathways implicated in Hodgkin lymphoma were identified. Selected proteins without previously demonstrated expression in Hodgkin lymphoma were validated by Western blot analysis (B-RAF, Erb-B3), immunofluorescence microscopy (Axin1, Tenascin-X, Mucin-2) and immunohistochemistry using a tissue microarray (BRAF, PIM1). This study represents the first comprehensive inventory of proteins expressed by Reed-Sternberg cells of Hodgkin lymphoma and demonstrates the utility of combining cellular subfractionation, protein precipitation, tandem mass spectrometry and bioinformatics analysis for comprehensive identification of proteins that may represent potential biomarkers of the disease.
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