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Mitochondrial proteomics of nasopharyngeal carcinoma metastasis.

Liu J, Zhan X, Li M, Li G, Zhang P, Xiao Z, Shao M, Peng F, Hu R, Chen Z - BMC Med Genomics (2012)

Bottom Line: The functional enrichment analyses of DEPs discovered five significant biological processes including cellular response to reactive oxygen species, hydrogen peroxide metabolic process, regulation of mitochondrial membrane potential, cell redox homeostasis and oxidation reduction, and five significant molecular functions including oxidoreductase activity, caspase inhibitor activity, peroxiredoxin activity, porin activity and antioxidant activity.Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential.These data suggest that those mitochondrial DEPs are potential biomarkers for NPC metastasis, and their dysregulation would play important roles in mitochondria oxidative stress-mediated NPC metastatic process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, Hunan, 410008, PR China.

ABSTRACT

Background: Mitochondrial proteomic alterations of nasopharyngeal carcinoma metastasis remain unknown. Our purpose is to screen mitochondrial proteins for the elucidation of the molecular mechanisms of nasopharyngeal carcinoma metastasis and the discovery of metastasis-related biomarkers.

Methods: Mitochondria were isolated from nasopharyngeal carcinoma metastatic (5-8F) and nonmetastatic (6-10B) cell lines, respectively. After characterization of isolated mitochondria, mitochondrial differentially expressed proteins (DEPs) were quantified by two-dimensional difference in-gel electrophoresis (2D-DIGE), and identified by peptide mass fingerprint (PMF) and tandem mass spectrometry (MS/MS). A functional enrichment analysis and a protein-protein interaction sub-network analysis for DEPs were carried out with bioinformatics. Furthermore, siRNAs transient transfections were used to suppress expressions of some up-regulated DEPs in metastatic cells (5-8F), followed by Transwell Migration assay.

Results: Sixteen mitochondrial DEPs including PRDX3 and SOD2 were identified. Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential. The functional enrichment analyses of DEPs discovered five significant biological processes including cellular response to reactive oxygen species, hydrogen peroxide metabolic process, regulation of mitochondrial membrane potential, cell redox homeostasis and oxidation reduction, and five significant molecular functions including oxidoreductase activity, caspase inhibitor activity, peroxiredoxin activity, porin activity and antioxidant activity. A protein-protein interaction sub-network of DEPs was generated with literature data. Ten mitochondrial DEPs including PRDX3, PRDX6, SOD2, ECH1, SERPINB5, COX5A, PDIA5, EIF5A, IDH3B, and PSMC4 were rationalized in the tumor-stroma co-evolution model that mitochondrial oxidative stress directly contributes to tumor metastasis.

Conclusions: Sixteen mitochondrial DEPs were identified with mass spectrometry and ten of them were rationalized in the tumor-stroma co-evolution model. Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential. These data suggest that those mitochondrial DEPs are potential biomarkers for NPC metastasis, and their dysregulation would play important roles in mitochondria oxidative stress-mediated NPC metastatic process.

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2D-DIGE analysis of metastatic 5-8F versus nonmetastatic 6-10B cells. A. A representative two-color merged 2D-DIGE gel image. Green is the Cy3-labeled metastatic 5-8F cell sample. Red is the Cy5-labeled 6-10B cell sample. B. Labeled differentially expressed proteins (DEPs) between metastatic (5-8F) and nonmetastatic (6-10B) cell mitochondria (Ratio = 5-8F/6-10B; Red: down-regulated; Green: up-regulated). C. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative up-regulated protein in metastatic 5-8F cell mitochondria (spot 2). D. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative down-regulated protein in metastatic 5-8F cell mitochondria (Spot 15).
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Figure 2: 2D-DIGE analysis of metastatic 5-8F versus nonmetastatic 6-10B cells. A. A representative two-color merged 2D-DIGE gel image. Green is the Cy3-labeled metastatic 5-8F cell sample. Red is the Cy5-labeled 6-10B cell sample. B. Labeled differentially expressed proteins (DEPs) between metastatic (5-8F) and nonmetastatic (6-10B) cell mitochondria (Ratio = 5-8F/6-10B; Red: down-regulated; Green: up-regulated). C. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative up-regulated protein in metastatic 5-8F cell mitochondria (spot 2). D. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative down-regulated protein in metastatic 5-8F cell mitochondria (Spot 15).

Mentions: Mitochondrial protein expressions were compared between metastatic 5-8F and nonmetastatic 6-10B cells using 2D-DIGE with a mixed-sample internal standard. After comparative analyses by DeCyder 5.0 software in all 9 protein-spot image maps, average of 1,047 ± 78 protein-spots were detected, in total 36 spots containing DEPs (X-fold5-8F/6-10B > 2, P < 0.05) were detected. Figure2A showed a representative two-color merged gel image. Figure2B showed 36 spots containing DEPs. Figure2C showed the three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of an up-regulated protein (spot 2). Figure2D showed the three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a down-regulated protein (spot 15).


Mitochondrial proteomics of nasopharyngeal carcinoma metastasis.

Liu J, Zhan X, Li M, Li G, Zhang P, Xiao Z, Shao M, Peng F, Hu R, Chen Z - BMC Med Genomics (2012)

2D-DIGE analysis of metastatic 5-8F versus nonmetastatic 6-10B cells. A. A representative two-color merged 2D-DIGE gel image. Green is the Cy3-labeled metastatic 5-8F cell sample. Red is the Cy5-labeled 6-10B cell sample. B. Labeled differentially expressed proteins (DEPs) between metastatic (5-8F) and nonmetastatic (6-10B) cell mitochondria (Ratio = 5-8F/6-10B; Red: down-regulated; Green: up-regulated). C. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative up-regulated protein in metastatic 5-8F cell mitochondria (spot 2). D. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative down-regulated protein in metastatic 5-8F cell mitochondria (Spot 15).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3539862&req=5

Figure 2: 2D-DIGE analysis of metastatic 5-8F versus nonmetastatic 6-10B cells. A. A representative two-color merged 2D-DIGE gel image. Green is the Cy3-labeled metastatic 5-8F cell sample. Red is the Cy5-labeled 6-10B cell sample. B. Labeled differentially expressed proteins (DEPs) between metastatic (5-8F) and nonmetastatic (6-10B) cell mitochondria (Ratio = 5-8F/6-10B; Red: down-regulated; Green: up-regulated). C. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative up-regulated protein in metastatic 5-8F cell mitochondria (spot 2). D. Three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a representative down-regulated protein in metastatic 5-8F cell mitochondria (Spot 15).
Mentions: Mitochondrial protein expressions were compared between metastatic 5-8F and nonmetastatic 6-10B cells using 2D-DIGE with a mixed-sample internal standard. After comparative analyses by DeCyder 5.0 software in all 9 protein-spot image maps, average of 1,047 ± 78 protein-spots were detected, in total 36 spots containing DEPs (X-fold5-8F/6-10B > 2, P < 0.05) were detected. Figure2A showed a representative two-color merged gel image. Figure2B showed 36 spots containing DEPs. Figure2C showed the three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of an up-regulated protein (spot 2). Figure2D showed the three-dimensional simulation, a close-up of the region of 2D-DIGE gel image, and the associated graph view of a down-regulated protein (spot 15).

Bottom Line: The functional enrichment analyses of DEPs discovered five significant biological processes including cellular response to reactive oxygen species, hydrogen peroxide metabolic process, regulation of mitochondrial membrane potential, cell redox homeostasis and oxidation reduction, and five significant molecular functions including oxidoreductase activity, caspase inhibitor activity, peroxiredoxin activity, porin activity and antioxidant activity.Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential.These data suggest that those mitochondrial DEPs are potential biomarkers for NPC metastasis, and their dysregulation would play important roles in mitochondria oxidative stress-mediated NPC metastatic process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, Hunan, 410008, PR China.

ABSTRACT

Background: Mitochondrial proteomic alterations of nasopharyngeal carcinoma metastasis remain unknown. Our purpose is to screen mitochondrial proteins for the elucidation of the molecular mechanisms of nasopharyngeal carcinoma metastasis and the discovery of metastasis-related biomarkers.

Methods: Mitochondria were isolated from nasopharyngeal carcinoma metastatic (5-8F) and nonmetastatic (6-10B) cell lines, respectively. After characterization of isolated mitochondria, mitochondrial differentially expressed proteins (DEPs) were quantified by two-dimensional difference in-gel electrophoresis (2D-DIGE), and identified by peptide mass fingerprint (PMF) and tandem mass spectrometry (MS/MS). A functional enrichment analysis and a protein-protein interaction sub-network analysis for DEPs were carried out with bioinformatics. Furthermore, siRNAs transient transfections were used to suppress expressions of some up-regulated DEPs in metastatic cells (5-8F), followed by Transwell Migration assay.

Results: Sixteen mitochondrial DEPs including PRDX3 and SOD2 were identified. Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential. The functional enrichment analyses of DEPs discovered five significant biological processes including cellular response to reactive oxygen species, hydrogen peroxide metabolic process, regulation of mitochondrial membrane potential, cell redox homeostasis and oxidation reduction, and five significant molecular functions including oxidoreductase activity, caspase inhibitor activity, peroxiredoxin activity, porin activity and antioxidant activity. A protein-protein interaction sub-network of DEPs was generated with literature data. Ten mitochondrial DEPs including PRDX3, PRDX6, SOD2, ECH1, SERPINB5, COX5A, PDIA5, EIF5A, IDH3B, and PSMC4 were rationalized in the tumor-stroma co-evolution model that mitochondrial oxidative stress directly contributes to tumor metastasis.

Conclusions: Sixteen mitochondrial DEPs were identified with mass spectrometry and ten of them were rationalized in the tumor-stroma co-evolution model. Those 5-8F cells with suppression of PRDX3 showed an increased mobility potential. These data suggest that those mitochondrial DEPs are potential biomarkers for NPC metastasis, and their dysregulation would play important roles in mitochondria oxidative stress-mediated NPC metastatic process.

Show MeSH
Related in: MedlinePlus