| 1. |
- Rahman-Roblick, Rubaiyat, et al.
(författare)
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p53 targets identified by protein expression profiling
- 2007
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 104:13, s. 5401-5406
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Tidskriftsartikel (refereegranskat)abstract
- p53 triggers cell cycle arrest and apoptosis through transcriptional regulation of specific target genes. We have investigated the effect of p53 activation on the proteome using 2D gel electrophoresis analysis of mitomycin C-treated HCT116 colon carcinoma cells carrying wild-type p53. Approximately 5,800 protein spots were separated in overlapping narrow-pH-range gel strips, and 115 protein spots showed significant expression changes upon p53 activation. The identity of 55 protein spots was obtained by mass spectrometry. The majority of the identified proteins have no previous connection to p53. The proteins fall into different functional categories, such as mRNA processing, translation, redox regulation, and apoptosis, consistent with the idea that p53 regulates multiple cellular pathways. p53-dependent regulation of five of the up-regulated proteins, eIF5A, hnRNP C1/C2, hnRNP K, lamin A/C, and Nm23-H1, and two of the down-regulated proteins, Prx II and TrpRS, was examined in further detail. Analysis of mRNA expression levels demonstrated both transcription-dependent and transcription-independent regulation among the identified targets. Thus, this study reveals protein targets of p53 and highlights the role of transcription-independent effects for the p53-induced biological response.
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| 2. |
- Gemoll, Timo, et al.
(författare)
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Chromosomal aneuploidy affects the global proteome equilibrium of colorectal cancer cells
- 2013
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Ingår i: Analytical Cellular Pathology. - 2210-7177 .- 2210-7185. ; 36:5-6, s. 149-161
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Chromosomal aneuploidy has been identified as a prognostic factor in the majority of sporadic carcinomas. However, it is not known how chromosomal aneuploidy affects chromosome-specific protein expression in particular, and the cellular proteome equilibrium in general. OBJECTIVE: The aim was to detect chromosomal aneuploidy-associated expression changes in cell clones carrying trisomies found in colorectal cancer. METHODS: We used microcell-mediated chromosomal transfer to generate three artificial trisomic cell clones of the karyotypically stable, diploid, yet mismatch-deficient, colorectal cancer cell line DLD1 - each of them harboring one extra copy of either chromosome 3, 7 or 13. Protein expression differences were assessed by two-dimensional gel electrophoresis and mass spectrometry, compared to whole-genome gene expression data, and evaluated by PANTHER classification system and Ingenuity Pathway Analysis (IPA). RESULTS: In total, 79 differentially expressed proteins were identified between the trisomic clones and the parental cell line. Up-regulation of PCNA and HMGB I as well as down-regulation of IDH3A and PSMB3 were revealed as trisomy-associated alterations involved in regulating genome stability. CONCLUSIONS: These results show that trisomies affect the expression of genes and proteins that are not necessarily located on the trisomic chromosome, but reflect a pathway-related alteration of the cellular equilibrium.
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| 3. |
- Gemoll, Timo, et al.
(författare)
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HDAC2 and TXNL1 distinguish aneuploid from diploid colorectal cancers
- 2011
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Ingår i: Cellular and Molecular Life Sciences (CMLS). - : Springer. - 1420-682X .- 1420-9071. ; 68:19, s. 3261-3274
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Tidskriftsartikel (refereegranskat)abstract
- DNA aneuploidy has been identified as a prognostic factor for epithelial malignancies. Further understanding of the translation of DNA aneuploidy into protein expression will help to define novel biomarkers to improve therapies and prognosis. DNA ploidy was assessed by image cytometry. Comparison of gel-electrophoresis-based protein expression patterns of three diploid and four aneuploid colorectal cancer cell lines detected 64 ploidy-associated proteins. Proteins were identified by mass spectrometry and subjected to Ingenuity Pathway Analysis resulting in two overlapping high-ranked networks maintaining Cellular Assembly and Organization, Cell Cycle, and Cellular Growth and Proliferation. CAPZA1, TXNL1, and HDAC2 were significantly validated by Western blotting in cell lines and the latter two showed expression differences also in clinical samples using a tissue microarray of normal mucosa (n=19), diploid (n=31), and aneuploid (n=47) carcinomas. The results suggest that distinct protein expression patterns, affecting TXNL1 and HDAC2, distinguish aneuploid with poor prognosis from diploid colorectal cancers.
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| 4. |
- Gemoll, Timo, et al.
(författare)
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Protein profiling of genomic instability in endometrial cancer
- 2012
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Ingår i: Cellular and Molecular Life Sciences (CMLS). - : Springer. - 1420-682X .- 1420-9071. ; 69:2, s. 325-333
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Tidskriftsartikel (refereegranskat)abstract
- DNA aneuploidy has been identified as a prognostic factor in the majority of epithelial malignancies. We aimed at identifying ploidy-associated protein expression in endometrial cancer of different prognostic subgroups. Comparison of gel electrophoresis-based protein expression patterns between normal endometrium (n = 5), diploid (n = 7), and aneuploid (n = 7) endometrial carcinoma detected 121 ploidy-associated protein forms, 42 differentially expressed between normal endometrium and diploid endometrioid carcinomas, 37 between diploid and aneuploid endometrioid carcinomas, and 41 between diploid endometrioid and aneuploid uterine papillary serous cancer. Proteins were identified by mass spectrometry and evaluated by Ingenuity Pathway Analysis. Targets were confirmed by liquid chromatography/mass spectrometry. Mass spectrometry identified 41 distinct polypeptides and pathway analysis resulted in high-ranked networks with vimentin and Nf-kappa B as central nodes. These results identify ploidy-associated protein expression differences that overrule histopathology-associated expression differences and emphasize particular protein networks in genomic stability of endometrial cancer.
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| 5. |
- Lundgren, Caroline, et al.
(författare)
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2-DE protein expression in endometrial carcinoma
- 2006
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Ingår i: Acta Oncologica. - Oslo : Taylor & Francis. - 0284-186X .- 1651-226X. ; 45:6, s. 685-694
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Tidskriftsartikel (refereegranskat)abstract
- The objective of this study was to explore the protein expression pattern in normal endometrial mucosa (n = 5) and endometrial carcinoma ( n = 15) of low ( diploid) and high ( aneuploid) malignancy potential by two-dimensional gel electrophoresis (2-DE). The specimens were evaluated for histopathologic subtype, stage and grade in relation to DNA ploidy. A match-set consisting of five samples from normal endometrium, eight diploid and seven aneuploid tumours was created. All the diploid and three of the aneuploid tumours were of endometrioid subtype, while the remaining four were of uterine seropapillary type. There were 192 protein spots differentiating diploid tumours from normal endometrium and 238 protein spots were separating aneuploid tumours from normal endometrium (p < 0.01). A cluster analysis based on 52 significantly deviating protein spots within the groups showed clustering and separation of the normal endometrium, diploid and aneuploid tumours. In conclusion this study showed significant differences in protein expression between normal endometrium and endometrial carcinoma as well as between endometrial carcinoma of low and high malignancy potential. In future studies these results may provide useful in finding new sensitive prognostic markers for endometrial cancer.
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| 6. |
- Roblick, Uwe Johannes
(författare)
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Colon cancer : disease related proteins in tumor tissue and serum
- 2004
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Despite the progress in surgical techniques and adjuvant therapies the mortality of around 60% within the group of colon cancer patients did not decrease significantly over the last decades. This determines a growing demand for biomarkers for early detection, prognosis and risk assessment in colorectal malignancies. Desirable criteria for such a biomarker test are minimal burden and maximum safety for the patient, cost efficiency and broad acceptance to reach a high compliance of the patients. More than 300 colon tissue samples and 1000 sera were obtained from CRC and FAP patients at the Lübeck and Düsseldorf University Hospitals in Germany. To identify the consequences of genetic aberrations on protein expression level, samples from patients with advanced sporadic colon cancers in which the corresponding mucosa, adenoma, carcinoma and liver metastasis were available, were analyzed by 2-D gel electrophoresis and mass spectrometry. A total of 46 proteins were found to be upregulated during the progression of sporadic cancer, and 26 were downregulated. Several of the identified polypeptides correlate with proteins regulating specific cell functions (cell cycle, cytoskeleton, metabolic pathways). In a further study we used a 2-DE based proteomic approach to compare the expression pattern of normal colonic mucosa vs. mucosa gained from FAP patients, polyps vs. FAP polyps and sporadic vs. FAP associated cancer, respectively. A total of 47 proteins were always present in FAP mucosa and absent in the normal mucosa. Based on a total of 37 proteins FAP polyps and sporadic polyps could be distinctly separated. In addition, the absence/presence pattern of 66 spots allowed to distinguish FAP cancers from sporadic cancers. These data suggest, that proteome analysis makes it possible to diagnose FAP already on macroscopically normal appearing colonic mucosa. By means of SELDI array based investigations we unrevealed 16 serum proteins that were able to classify 98% of all test set samples correctly. Our SELDI results show that serum marker protein profiling enables to diagnose and discern malignant colon cancer patients from healthy individuals. Although these markers need validation before they can be used in clinics they have potential for the design of a marker panel for objective diagnosis and therapeutic strategies for colorectal cancer and metastasis.
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| 7. |
- Roblick, Uwe J., et al.
(författare)
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Proteomic analysis of protein expression in human tonsillar cancer - differentially expressed proteins characterize human tonsillar cancer
- 2008
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Ingår i: Acta Oncologica. - : Informa UK Limited. - 0284-186X .- 1651-226X. ; 47:8, s. 1493-1501
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Head and neck cancer continues to be one of the most common tumor entities worldwide. Within this group of malignancies, tonsillar squamous cell carcinoma represent approximately 15-20% of all intraoral and oropharyngeal carcinomas in the United States. Accurate and early stage diagnosis still remains a major challenge, as patients are often presented at an advanced stage of disease, causing a low overall survival rate. Thus, new diagnostic markers are highly desirable and could allow for a more reliable diagnosis, with further insights into carcinogenesis and tumor biology. Furthermore, these markers could be the basis for new therapeutic targets and early disease detection. To address these issues, we decided to use a global proteomic approach to characterize tonsillar squamous cell carcinoma. MATERIALS AND METHODS: A total of 19 tonsillar carcinoma samples and 12 benign controls acquired from the corresponding normal epithelium were analyzed by 2-D gel electrophoresis. 2-DE gels were silver stained and analyzed using the PDQuest analysis software (BioRad). Tumor specific spots were detected and identified by consecutive MALDI-TOF-MS or MS/MS polypeptide identification. RESULTS: In total, 70 proteins showed significant quantitative differences in protein expression, with 50 polypeptides accessible for identification. Of those 50 polypeptides, we were able to identify a total of 27 proteins and protein isoforms, significantly up- or down-regulated in tonsillar cancer samples. In addition to previously reported polypeptides in head and neck cancers, we were able to identify several new potential marker proteins in this study. CONCLUSION: Our results show that a combination of tonsillar cancer specific proteins can be used for histopathological diagnosis and may serve as a basis for discovering further biomarkers for early detection and prediction of response to treatment in the future.
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