Towards the Application of Proteomics to Human Thyroid Tissue

Krause, Kerstin; Schierhorn, Angelika; Sinz, Andrea; Wissmann, Johannes-Dieter; Beck‐Sickinger, Annette G.; Paschke, Ralf; Führer, Dagmar

doi:10.1089/thy.2006.16.1131

Cited by 17 publications

(13 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…15 Thyroglobulin measurement was performed using the Brahms thyroglobulin RIA-Kit (BRAHMS AG, Hennigsdorf, Germany), and sample load for protein expression analysis was corrected for thyroglobulin content as previously described. 15 Isoelectric focusing was performed in 24-cm-long, pH 4 to 7 immobilized pH gradient (IPG) strips (GE Healthcare, Freiburg, Germany) followed by SDS-PAGE in 8% to 16% gradient gels. 15 All samples were run in triplicate.…”

Section: Sample Preparation Protein Extraction and Two-dimensional mentioning

confidence: 99%

“…15 Isoelectric focusing was performed in 24-cm-long, pH 4 to 7 immobilized pH gradient (IPG) strips (GE Healthcare, Freiburg, Germany) followed by SDS-PAGE in 8% to 16% gradient gels. 15 All samples were run in triplicate.…”

Section: Sample Preparation Protein Extraction and Two-dimensional mentioning

confidence: 99%

“…For spot picking, a preparative gel with a 1000-g protein load was run and stained with colloidal Coomassie blue after scanning. 15 …”

Section: Protein Detection and Spot Analysismentioning

confidence: 99%

“…20 To avoid the influence of different thyroglobulin protein contents on the protein patterns of the respective samples, we adapted the protein loads per gel to the thyroglobulin content of the sample. 15 All samples that had been analyzed by genotyping were subjected to this proteomic approach.…”

Section: Differential Protein Expression In Fa and Ftcmentioning

confidence: 99%

See 3 more Smart Citations

Dissecting Molecular Events in Thyroid Neoplasia Provides Evidence for Distinct Evolution of Follicular Thyroid Adenoma and Carcinoma

Krause¹,

Prawitt²,

Eszlinger³

et al. 2011

The American Journal of Pathology

Self Cite

View full text Add to dashboard Cite

Benign hypofunctional cold thyroid nodules (CTNs) are a frequent scintiscan finding and need to be distinguished from thyroid carcinomas. The origin of CTNs with follicular morphologic features is unresolved. The DNA damage response might act as a physiologic barrier, inhibiting the progression of preneoplastic lesions to neoplasia. We investigated the following in hypofunctional follicular adenoma (FA) and follicular thyroid cancer (FTC): i) the mutation rate of frequently activated oncogenes, ii) the activation of DNA damage response checkpoints, and iii) the differential proteomic pattern between FA and FTC. Both FTC and FA, which did not harbor RAS, phosphoinositide-3-kinase, or PAX/peroxisome proliferator activated receptor-␥ mutations, express various proteins in common and others that are more distinctly expressed in FTC rather than in FA or normal thyroid tissue. This finding is in line with the finding of constitutive DNA damage checkpoint activation (p-Chk2, ␥-H2AX) and evidence for replicative stress causing genomic instability (increased cyclin E, retinoblastoma, or E2F1 mRNA expression) in FTC but not FA. We discuss the findings of the increased expression of translationally controlled tumor protein, phosphatase 2A inhibitor, and DJ-1 in FTC compared with FA identified by proteomics and their potential implication in follicular thyroid carcinogenesis. Our present findings argue for the definition of FA as a truly benign entity and against progressive development of FA to FTC.

show abstract

Section: Sample Preparation Protein Extraction and Two-dimensional mentioning

confidence: 99%

Section: Sample Preparation Protein Extraction and Two-dimensional mentioning

confidence: 99%

“…For spot picking, a preparative gel with a 1000-g protein load was run and stained with colloidal Coomassie blue after scanning. 15 …”

Section: Protein Detection and Spot Analysismentioning

confidence: 99%

Section: Differential Protein Expression In Fa and Ftcmentioning

confidence: 99%

See 2 more Smart Citations

Dissecting Molecular Events in Thyroid Neoplasia Provides Evidence for Distinct Evolution of Follicular Thyroid Adenoma and Carcinoma

Krause¹,

Prawitt²,

Eszlinger³

et al. 2011

The American Journal of Pathology

Self Cite

View full text Add to dashboard Cite

show abstract

“…These proteins were more abundant in PTC and differed in posttranslational modifications (glycosylation), which would not have been detected by genomic approaches. Proteomic analyses have also been carried out in human thyroid carcinoma cell lines (8-10), FRTL-5 cells (11), human tumor tissues (3,9,12,13), and fine-needle aspiration biopsies (14,15).…”

mentioning

confidence: 99%

Applications of Proteomics to Thyroid Neoplasms: Are We There Yet?

Haugen¹,

Duncan²

2010

Thyroid

View full text Add to dashboard Cite

Application of free‐flow IEF to identify protein candidates changing under microgravity conditions

et al. 2010

View full text Add to dashboard Cite

Using antibody-related methods, we recently found that human thyroid cells express various proteins differently depending on whether they are cultured under normal gravity (1g) or simulated microgravity (s-mg). In this study, we performed proteome analysis in order to identify more gravity-sensitive thyroid proteins. Cells cultured under 1g or s-mg conditions were sonicated. Proteins released into the supernatant and those remaining in the cell fragments were fractionated by free-flow IEF. The fractions obtained were further separated by SDS-gel electrophoresis. Selected gel pieces were excised and their proteins were determined by MS. A total of 235 different proteins were found. Out of 235 proteins, 37 appeared to be first identifications in human thyroid cells. Comparing SDS gel lanes of equally numbered free-flow IEF fractions revealed similar patterns with a number of identical bands if proteins of a distinct cell line had been applied, irrespective of whether the cells had been cultured under 1g or s-mg. Most of the identical band pairs contained identical proteins. However, the concentrations of some types of proteins were different within the two pieces of gel. Proteins that concentrated differently in such pieces of gel are considered as candidates for further investigations of gravitational sensitivity.

show abstract

Towards the Application of Proteomics to Human Thyroid Tissue

Abstract: Application of the described methods provides the basis for a highly sensitive and reproducible proteome analysis of the human thyroid, providing an additional novel tool to elucidate complex proteins changes in human thyroid biology as well as pathophysiology of human thyroid disease.

Cited by 17 publications

References 49 publications

Dissecting Molecular Events in Thyroid Neoplasia Provides Evidence for Distinct Evolution of Follicular Thyroid Adenoma and Carcinoma

Dissecting Molecular Events in Thyroid Neoplasia Provides Evidence for Distinct Evolution of Follicular Thyroid Adenoma and Carcinoma

Applications of Proteomics to Thyroid Neoplasms: Are We There Yet?

Application of free‐flow IEF to identify protein candidates changing under microgravity conditions

Contact Info

Product

Resources

About