Immunoassay-based proteome profiling of 24 pancreatic cancer cell lines

Alhamdani, Mohamed Saiel Saeed; Youns, Mаhmoud; Buchholz, Malte; Gress, Thomas M.; Beckers, Marie-Claire; Maréchal, Daniel; Bauer, Andrea S.; Schröder, Christoph; Hoheisel, Jörg D.

doi:10.1016/j.jprot.2012.04.042

Cited by 27 publications

(36 citation statements)

References 78 publications

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“…The same format was used for a recent comparative analysis of 24 pancreatic cancer cell lines (28). All measurements were done on five biological replicates for each condition.…”

Section: Resultsmentioning

confidence: 99%

“…As compared with measurements with TNF-␣ and CCL4, PSC growth in presence of IL6 exhibited reduced cellular movement, required for migration and invasion, but still higher values than those measured in serum-free medium. Significantly elevated levels of IL6 have been reported in pancreatic cancer cell lines (28) and medium of these cells (56) as well as in sera of pancreatic cancer patients (57). The anti-apoptotic effect of IL6 is well documented (58,59).…”

Section: Discussionmentioning

confidence: 99%

“…In the analysis, a set of 810 antibodies was used, as reported earlier (28). Their target proteins and origin are listed in supplemental Table S1.…”

Section: Methodsmentioning

confidence: 99%

“…Also, 35 g of a common reference sample was added to all incubations and labeled with the second dye (33). The reference sample was made by pooling protein extracts prepared from 24 pancreatic cancer cell lines (28). Aliquots were stored frozen until use.…”

Section: Methodsmentioning

confidence: 99%

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Proteome Variations in Pancreatic Stellate Cells upon Stimulation with Proinflammatory Factors

Marzoq

Giese

Hoheisel

et al. 2013

Journal of Biological Chemistry

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Background: Stroma formation in pancreatic cancer is controlled via the activation of pancreatic stellate cells. Results: Pancreatic stellate cells demonstrate different proteomic and functional attributes following specific stimuli. Conclusion: The proteomic feature of pancreatic stellate cells shows TNF-␣ as a main contributor to activation. Significance: The shift of proteomes in activated cells may help in understanding the processes of stromal formation and finding new drug targets.

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“…The same format was used for a recent comparative analysis of 24 pancreatic cancer cell lines (28). All measurements were done on five biological replicates for each condition.…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…In the analysis, a set of 810 antibodies was used, as reported earlier (28). Their target proteins and origin are listed in supplemental Table S1.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Proteome Variations in Pancreatic Stellate Cells upon Stimulation with Proinflammatory Factors

Marzoq

Giese

Hoheisel

et al. 2013

Journal of Biological Chemistry

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“…It is also well established that human tumors are highly heterogeneous both intra-and interindividually. Given the fact that the majority of established long-term passaged cancer cell lines do not reflect either complexity, or individual features of the parental tumors [3], [38], it is conceivable that modern cancer research and anti-cancer drug development should be based on novel in vitro and in vivo systems allowing for physiologically and clinically more relevant modeling and more objective evaluation of drug efficacy. Molecular, biochemical, genomic and proteomic characterization of human tumors aims to discover novel targets that potentially can cure cancer.…”

Section: Discussionmentioning

confidence: 99%

Establishment of Highly Tumorigenic Human Colorectal Cancer Cell Line (CR4) with Properties of Putative Cancer Stem Cells

et al. 2014

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BackgroundColorectal cancer (CRC) has the third highest mortality rates among the US population. According to the most recent concept of carcinogenesis, human tumors are organized hierarchically, and the top of it is occupied by malignant stem cells (cancer stem cells, CSCs, or cancer-initiating cells, CICs), which possess unlimited self-renewal and tumor-initiating capacities and high resistance to conventional therapies. To reflect the complexity and diversity of human tumors and to provide clinically and physiologically relevant cancer models, large banks of characterized patient-derived low-passage cell lines, and especially CIC-enriched cell lines, are urgently needed.Principal FindingsHere we report the establishment of a novel CIC-enriched, highly tumorigenic and clonogenic colon cancer cell line, CR4, derived from liver metastasis. This stable cell line was established by combining 3D culturing and 2D culturing in stem cell media, subcloning of cells with particular morphology, co-culture with carcinoma associated fibroblasts (CAFs) and serial transplantation to NOD/SCID mice. Using RNA-Seq complete transcriptome profiling of the tumorigenic fraction of the CR4 cells in comparison to the bulk tumor cells, we have identified about 360 differentially expressed transcripts, many of which represent stemness, pluripotency and resistance to treatment. Majority of the established CR4 cells express common markers of stemness, including CD133, CD44, CD166, EpCAM, CD24 and Lgr5. Using immunocytochemical, FACS and western blot analyses, we have shown that a significant ratio of the CR4 cells express key markers of pluripotency markers, including Sox-2, Oct3/4 and c-Myc. Constitutive overactivation of ABC transporters and NF-kB and absence of tumor suppressors p53 and p21 may partially explain exceptional drug resistance of the CR4 cells.ConclusionsThe highly tumorigenic and clonogenic CIC-enriched CR4 cell line may provide an important new tool to support the discovery of novel diagnostic and/or prognostic biomarkers as well as the development of more effective therapeutic strategies.

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Antibody Microarrays

Wingren

Borrebaeck

2015

Encyclopedia of Life Sciences

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Antibody‐based microarray is a recently established proteomic technology, providing unique opportunities for multiplexed protein expression profiling of crude, non‐fractionated samples in a specific, sensitive and miniaturised manner. A microarray can be printed with a few up to several thousands of different antibodies carrying the desired specificities, the biological sample added (e.g. a serum sample) and any specifically bound analytes detected and quantified. The obtained microarray images are then transformed into protein expression profiles, or protein maps, revealing the composition of the sample at the molecular level. Bioinformatics is used to handle the often large‐scale data set(s) acquired. Protein expression profiling using this tool will provide new opportunities for biomarker discovery, drug target identification, (early) disease diagnostics and prognostics as well as insights into disease biology. Key Concepts Antibody microarrays are based on dispensing antibodies one‐by‐one in an ordered pattern, an array, onto a solid support. Antibody‐based microarray is a miniaturised set‐up for simultaneously measuring numerous proteins in minute amount of sample. Antibody microarrays can be used to perform multiplexed protein expression profiling of crude, complex samples. Using antibody microarrays, less than a drop of blood is required in order to generate a protein map, or protein fingerprint, reflecting the protein content in the sample. Multiplexed protein expression profiling using antibody microarrays provides novel opportunities for biomarker discovery, disease diagnosis and prognosis.

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Immunoassay-based proteome profiling of 24 pancreatic cancer cell lines

Cited by 27 publications

References 78 publications

Proteome Variations in Pancreatic Stellate Cells upon Stimulation with Proinflammatory Factors

Proteome Variations in Pancreatic Stellate Cells upon Stimulation with Proinflammatory Factors

Establishment of Highly Tumorigenic Human Colorectal Cancer Cell Line (CR4) with Properties of Putative Cancer Stem Cells

Antibody Microarrays

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