Limits...
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.

Show MeSH

Related in: MedlinePlus

Effects of PRDX3 siRNA transient transfection on the migration capability of metastatic 5-8F cells. Effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells. The top shows four representative micrographs of metastatic 5-8F cells migrated through Transwell insert membranes among four experimental groups (400 ×). The bottom shows the quantitative comparison of the number of cells that migrated through Transwell insert membranes. Group A (a): nonmetastatic 6-10B cells. Group B (b): metastatic 5-8F cells. Group C (c): metastatic 5-8F cells with PRDX3 siRNA transient transfection. Group D (d): metastatic 5-8F cells with a scramble sequence siRNA transient transfection. Transwell migration assays were carried out triply (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3539862&req=5

Figure 8: Effects of PRDX3 siRNA transient transfection on the migration capability of metastatic 5-8F cells. Effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells. The top shows four representative micrographs of metastatic 5-8F cells migrated through Transwell insert membranes among four experimental groups (400 ×). The bottom shows the quantitative comparison of the number of cells that migrated through Transwell insert membranes. Group A (a): nonmetastatic 6-10B cells. Group B (b): metastatic 5-8F cells. Group C (c): metastatic 5-8F cells with PRDX3 siRNA transient transfection. Group D (d): metastatic 5-8F cells with a scramble sequence siRNA transient transfection. Transwell migration assays were carried out triply (n = 3).

Mentions: In three independent experiments, the number of cells migrated through Transwell insert membrane was counted for four experimental groups: Group A (nonmetastatic 6-10B cells; 0 ± 0), Group B (metastatic 5-8F cells; 647 ± 35), Group C (metastatic 5-8F cells with PRDX3 siRNA transient transfection; 838 ± 31), and Group D (metastatic 5-8F cells with a scramble sequence siRNA transient transfection; 593 ± 27). The number of Group C was about 40% more than the number of Group B and Group D (P < 0.05). The effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells was showed (Figure8). It demonstrates that suppression of PRDX3 in metastatic 5-8F cells increased its mobility potential.


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)

Effects of PRDX3 siRNA transient transfection on the migration capability of metastatic 5-8F cells. Effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells. The top shows four representative micrographs of metastatic 5-8F cells migrated through Transwell insert membranes among four experimental groups (400 ×). The bottom shows the quantitative comparison of the number of cells that migrated through Transwell insert membranes. Group A (a): nonmetastatic 6-10B cells. Group B (b): metastatic 5-8F cells. Group C (c): metastatic 5-8F cells with PRDX3 siRNA transient transfection. Group D (d): metastatic 5-8F cells with a scramble sequence siRNA transient transfection. Transwell migration assays were carried out triply (n = 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 8: Effects of PRDX3 siRNA transient transfection on the migration capability of metastatic 5-8F cells. Effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells. The top shows four representative micrographs of metastatic 5-8F cells migrated through Transwell insert membranes among four experimental groups (400 ×). The bottom shows the quantitative comparison of the number of cells that migrated through Transwell insert membranes. Group A (a): nonmetastatic 6-10B cells. Group B (b): metastatic 5-8F cells. Group C (c): metastatic 5-8F cells with PRDX3 siRNA transient transfection. Group D (d): metastatic 5-8F cells with a scramble sequence siRNA transient transfection. Transwell migration assays were carried out triply (n = 3).
Mentions: In three independent experiments, the number of cells migrated through Transwell insert membrane was counted for four experimental groups: Group A (nonmetastatic 6-10B cells; 0 ± 0), Group B (metastatic 5-8F cells; 647 ± 35), Group C (metastatic 5-8F cells with PRDX3 siRNA transient transfection; 838 ± 31), and Group D (metastatic 5-8F cells with a scramble sequence siRNA transient transfection; 593 ± 27). The number of Group C was about 40% more than the number of Group B and Group D (P < 0.05). The effect of PRDX3 siRNA transient transfection on migration capability of metastatic 5-8F cells was showed (Figure8). It demonstrates that suppression of PRDX3 in metastatic 5-8F cells increased its mobility potential.

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