Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme

Timothy W. Vogel, Zhengping Zhuang, Jie Li, Hiroaki Okamoto, Makoto Furuta, Youn Soo Lee, Weifen Zeng, Edward H. Oldfield, Alexander Vortmeyer, Robert J. Weil

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Introduction: Research into the pathogenesis, molecular signaling, and treatment of glioblastoma multiforme (GBM) has traditionally been conducted using cell lines derived from malignant gliomas. We compared protein expression patterns between solid primary GBMs and GBM cell lines to identify proteins whose expression may be altered in cell culture. Methods: We cultured cell lines U87, U118, U251, and A172 and used tissue-selective microdissection of eight primary GBMs to obtain pure populations of tumor cells, which we studied using two-dimensional gel electrophoresis (2DGE) and examined using differential expression software. Select protein targets expressed differentially between GBM tumors and GBM cell lines were sequenced using tandem mass spectrometry. Results: Analysis of the primary GBM tumor samples (n = 8) and the GBM cell lines revealed reproducibly similar proteomic patterns for each group, which distinguished tumors from the cell lines. Gels contained up to 500 proteins that were consistently identified in the pH 4 to 7 range. Comparison of proteins identified in the GBM tumors and in the cell lines showed ∼160 proteins that were gained and 60 proteins that were lost on culture. Using normalized intensity patterns from the 2DGE images, ANOVA tests were done and statistically significant spots were identified. Seven proteins found in the cell lines were significantly increased when compared with the GBM tumors (P < 0.05), whereas 10 proteins were significantly decreased from cell lines compared with the GBM tumors. Proteins identified included transcription factors, tumor suppressor genes, cytoskeletal proteins, and cellular metabolic proteins. Conclusion: Global protein and proteomic differences were identified between primary GBM tumor samples and GBM cell lines. The proteins identified by 2DGE analysis elucidate some of the selection pressures of in vitro culture, help accentuate the advantages and limitations of cell culture, and may aid comprehension of gliomagenesis and enhance development of new therapeutics.

Original languageEnglish (US)
Pages (from-to)3624-3632
Number of pages9
JournalClinical Cancer Research
Volume11
Issue number10
DOIs
StatePublished - May 15 2005
Externally publishedYes

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Glioblastoma
Glioma
Cell Line
Proteins
Neoplasms
Tumor Cell Line
Proteomics
Cell Culture Techniques
Microdissection
Cytoskeletal Proteins
Electrophoresis, Gel, Two-Dimensional
Tandem Mass Spectrometry
Tumor Suppressor Genes
Cultured Cells
Analysis of Variance
Transcription Factors
Software
Gels

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Vogel, T. W., Zhuang, Z., Li, J., Okamoto, H., Furuta, M., Lee, Y. S., ... Weil, R. J. (2005). Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme. Clinical Cancer Research, 11(10), 3624-3632. https://doi.org/10.1158/1078-0432.CCR-04-2115

Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme. / Vogel, Timothy W.; Zhuang, Zhengping; Li, Jie; Okamoto, Hiroaki; Furuta, Makoto; Lee, Youn Soo; Zeng, Weifen; Oldfield, Edward H.; Vortmeyer, Alexander; Weil, Robert J.

In: Clinical Cancer Research, Vol. 11, No. 10, 15.05.2005, p. 3624-3632.

Research output: Contribution to journalArticle

Vogel, TW, Zhuang, Z, Li, J, Okamoto, H, Furuta, M, Lee, YS, Zeng, W, Oldfield, EH, Vortmeyer, A & Weil, RJ 2005, 'Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme', Clinical Cancer Research, vol. 11, no. 10, pp. 3624-3632. https://doi.org/10.1158/1078-0432.CCR-04-2115
Vogel, Timothy W. ; Zhuang, Zhengping ; Li, Jie ; Okamoto, Hiroaki ; Furuta, Makoto ; Lee, Youn Soo ; Zeng, Weifen ; Oldfield, Edward H. ; Vortmeyer, Alexander ; Weil, Robert J. / Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme. In: Clinical Cancer Research. 2005 ; Vol. 11, No. 10. pp. 3624-3632.
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T1 - Proteins and protein pattern differences between glioma cell lines and glioblastoma multiforme

AU - Vogel, Timothy W.

AU - Zhuang, Zhengping

AU - Li, Jie

AU - Okamoto, Hiroaki

AU - Furuta, Makoto

AU - Lee, Youn Soo

AU - Zeng, Weifen

AU - Oldfield, Edward H.

AU - Vortmeyer, Alexander

AU - Weil, Robert J.

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N2 - Introduction: Research into the pathogenesis, molecular signaling, and treatment of glioblastoma multiforme (GBM) has traditionally been conducted using cell lines derived from malignant gliomas. We compared protein expression patterns between solid primary GBMs and GBM cell lines to identify proteins whose expression may be altered in cell culture. Methods: We cultured cell lines U87, U118, U251, and A172 and used tissue-selective microdissection of eight primary GBMs to obtain pure populations of tumor cells, which we studied using two-dimensional gel electrophoresis (2DGE) and examined using differential expression software. Select protein targets expressed differentially between GBM tumors and GBM cell lines were sequenced using tandem mass spectrometry. Results: Analysis of the primary GBM tumor samples (n = 8) and the GBM cell lines revealed reproducibly similar proteomic patterns for each group, which distinguished tumors from the cell lines. Gels contained up to 500 proteins that were consistently identified in the pH 4 to 7 range. Comparison of proteins identified in the GBM tumors and in the cell lines showed ∼160 proteins that were gained and 60 proteins that were lost on culture. Using normalized intensity patterns from the 2DGE images, ANOVA tests were done and statistically significant spots were identified. Seven proteins found in the cell lines were significantly increased when compared with the GBM tumors (P < 0.05), whereas 10 proteins were significantly decreased from cell lines compared with the GBM tumors. Proteins identified included transcription factors, tumor suppressor genes, cytoskeletal proteins, and cellular metabolic proteins. Conclusion: Global protein and proteomic differences were identified between primary GBM tumor samples and GBM cell lines. The proteins identified by 2DGE analysis elucidate some of the selection pressures of in vitro culture, help accentuate the advantages and limitations of cell culture, and may aid comprehension of gliomagenesis and enhance development of new therapeutics.

AB - Introduction: Research into the pathogenesis, molecular signaling, and treatment of glioblastoma multiforme (GBM) has traditionally been conducted using cell lines derived from malignant gliomas. We compared protein expression patterns between solid primary GBMs and GBM cell lines to identify proteins whose expression may be altered in cell culture. Methods: We cultured cell lines U87, U118, U251, and A172 and used tissue-selective microdissection of eight primary GBMs to obtain pure populations of tumor cells, which we studied using two-dimensional gel electrophoresis (2DGE) and examined using differential expression software. Select protein targets expressed differentially between GBM tumors and GBM cell lines were sequenced using tandem mass spectrometry. Results: Analysis of the primary GBM tumor samples (n = 8) and the GBM cell lines revealed reproducibly similar proteomic patterns for each group, which distinguished tumors from the cell lines. Gels contained up to 500 proteins that were consistently identified in the pH 4 to 7 range. Comparison of proteins identified in the GBM tumors and in the cell lines showed ∼160 proteins that were gained and 60 proteins that were lost on culture. Using normalized intensity patterns from the 2DGE images, ANOVA tests were done and statistically significant spots were identified. Seven proteins found in the cell lines were significantly increased when compared with the GBM tumors (P < 0.05), whereas 10 proteins were significantly decreased from cell lines compared with the GBM tumors. Proteins identified included transcription factors, tumor suppressor genes, cytoskeletal proteins, and cellular metabolic proteins. Conclusion: Global protein and proteomic differences were identified between primary GBM tumor samples and GBM cell lines. The proteins identified by 2DGE analysis elucidate some of the selection pressures of in vitro culture, help accentuate the advantages and limitations of cell culture, and may aid comprehension of gliomagenesis and enhance development of new therapeutics.

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