Proteins and Protein Pattern Differences between Glioma Cell Lines and Glioblastoma Multiforme

Abstract

Introduction: Researchinto 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 f160 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.

Protein profiling of formalin fixed paraffin embedded tissue: Identification of potential biomarkers for pediatric brainstem glioma

“…Our data agree with previous microarray gene expression profiling studies that have used either brain tumor cell lines or tissue from adult supratentorial gliomas [28][29][30]. We found that 15 out of the 54 differentially expressed proteins have been reported in these previous studies as potential tumor biomarkers.…”

“…We found that 15 out of the 54 differentially expressed proteins have been reported in these previous studies as potential tumor biomarkers. Among these, vimentin, peroxiredoxin-6, SOD1, annexin V, GFAP, and DRP-2 have been shown to be up-regulated in adult glioblastoma multiform (GBM) [28][29][30][31] whereas tubulin beta-2C chain, neurofilament light polypeptide, and pyruvate kinase are known to be up-regulated in meduloblastoma cell lines [29,32]. We also detected a high expression of angiogenesis-related proteins in BSG tumor compared to the normal tissue sections and these included hemoglobin subunits, which are an indication of angiogenesis, as well as periostin and mimecan precursors.…”

Altered expression of alpha-dystroglycan subunit in human gliomas

“…A vast number of reports deal with the biological differences between the U87MG and Dystroglycan in Human Gliomas the U251MG cell lines both in vitro and in vivo. 37,38 Xenograft implantations of U87MG and U251MG cells into the brain of nude mice display consistent and reproducible differences in tumor growth and invasion, which could also reflect the differences in α-DG glycosylation and in their proliferative response to DG overexpression. 39 In fact, the α-DG negative U87MG are more tumorigenic in nude mice brain, 40 and they grow mainly as expanding tumor mass by making spheres with little perivascular infiltration.…”

Development of barcode and proteome profiling of glioblastoma

“…Следует заметить, что за последние годы протеомный сравнительный анализ в области изучения глиобластомы стал очень популярен, поэтому есть возможность для сравнения наших и уже опубликованных результатов. Необходимо отметить, что по многим белкам наши и литературные данные совпадают, например, по пируваткиназе М1/М2 [24], триозофосфатизомеразе [25], кофилину, енолазе, виментину и его разным формам [26,27]. Появление мутантных форм р53 характерно для многих опухолей, есть публикации такого рода и по глиобластомам [28].…”