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

Experimental data-based diagram that rationalizes differentially expressed mitochondrial proteins in a tumor-stroma co-evolution model. ↑: up-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. ↓: down-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. The green frame means the relationship between proteins and tumor metastasis is unclear. The blue frame means the relationship between proteins and tumor metastasis is, at least partially, clear. The blue frame with solid line means protein promotes metastasis and the blue frame with dotted line means protein inhibits metastasis. SOD2, superoxide dismutase [Mn] mitochondrial isoform B; PRDX3, peroxiredoxin 3; PRDX6, peroxiredoxin 6; ECH1, mitochondrial delta(3,5)-delta(2,4)-dienoyl-CoA isomerase; SERPINB5, maspin; COX5A, mitochondrial cytochrome c oxidase subunit 5A; PDIA5, protein disulfide; EIF5A, eukaryotic initiation factor 5A; IDH3B, mitochondrial isocitrate dehydrogenase [NAD] subunit alpha; ETFA, electron transfer flavoprotein; PHB, prohibitin; MRPL12, mitochondrial ribosomal protein L7/L12; PSMC4, 26S protease regulatory subunit 7 isoform 1; VDAC1 and VDAC2, voltage-dependent anion channel 1 and 2; and CALU, calumenin isoform c precursor.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: Experimental data-based diagram that rationalizes differentially expressed mitochondrial proteins in a tumor-stroma co-evolution model. ↑: up-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. ↓: down-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. The green frame means the relationship between proteins and tumor metastasis is unclear. The blue frame means the relationship between proteins and tumor metastasis is, at least partially, clear. The blue frame with solid line means protein promotes metastasis and the blue frame with dotted line means protein inhibits metastasis. SOD2, superoxide dismutase [Mn] mitochondrial isoform B; PRDX3, peroxiredoxin 3; PRDX6, peroxiredoxin 6; ECH1, mitochondrial delta(3,5)-delta(2,4)-dienoyl-CoA isomerase; SERPINB5, maspin; COX5A, mitochondrial cytochrome c oxidase subunit 5A; PDIA5, protein disulfide; EIF5A, eukaryotic initiation factor 5A; IDH3B, mitochondrial isocitrate dehydrogenase [NAD] subunit alpha; ETFA, electron transfer flavoprotein; PHB, prohibitin; MRPL12, mitochondrial ribosomal protein L7/L12; PSMC4, 26S protease regulatory subunit 7 isoform 1; VDAC1 and VDAC2, voltage-dependent anion channel 1 and 2; and CALU, calumenin isoform c precursor.

Mentions: Each mitochondrial DEP was rationalized in the tumor-stroma co-evolution model, and an experimental data-based diagram was proposed (Figure9). In our comparative proteomic study of mitochondria between NPC metastatic 5-8F and nonmetastatic 6-10B cells, the first five enriched GO terms of DEPs were closely related to oxidative stress (Table2). For PRDX3, our comparative proteomic data demonstrated the expression of PRDX3 in mitochondria of metastatic 5-8F cells is higher than in mitochondria of nonmetastatic 6-10B cells, it shows that the anti-oxidative stress capability in metastatic 5-8F cells is stronger than nonmetastatic 6-10B cells. This result is consistent with previous conclusion[14] that the enhanced anti-oxidant capabilities in cancer cells defend themselves from excess stromal oxidative damage and favors cell metastasis. Furthermore, among several DEPs (data not showed) selected to carry out siRNA transient transfections and then Transwell Migration assay, only suppression of PRDX3 expression resulted in the significantly increased number of cells migrated through Transwell membrane (P < 0.05). PRDX3, mitochondria thioredoxin-dependent peroxide reductase, is the main way by which cancer cells reduce their levels of H2O2 built up during active respiration[17,18], suppression of this enzyme, a scavenger of peroxide, could increase oxidative stress in cancer cells to up-regulate the redox-signaling, promote the glycolysis, proliferation, and survival, and finally lead increased cells mobility potential[14] (Figure8). To our knowledge, this was the first experimental evidence that PRDX3 was related to NPC metastasis.


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)

Experimental data-based diagram that rationalizes differentially expressed mitochondrial proteins in a tumor-stroma co-evolution model. ↑: up-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. ↓: down-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. The green frame means the relationship between proteins and tumor metastasis is unclear. The blue frame means the relationship between proteins and tumor metastasis is, at least partially, clear. The blue frame with solid line means protein promotes metastasis and the blue frame with dotted line means protein inhibits metastasis. SOD2, superoxide dismutase [Mn] mitochondrial isoform B; PRDX3, peroxiredoxin 3; PRDX6, peroxiredoxin 6; ECH1, mitochondrial delta(3,5)-delta(2,4)-dienoyl-CoA isomerase; SERPINB5, maspin; COX5A, mitochondrial cytochrome c oxidase subunit 5A; PDIA5, protein disulfide; EIF5A, eukaryotic initiation factor 5A; IDH3B, mitochondrial isocitrate dehydrogenase [NAD] subunit alpha; ETFA, electron transfer flavoprotein; PHB, prohibitin; MRPL12, mitochondrial ribosomal protein L7/L12; PSMC4, 26S protease regulatory subunit 7 isoform 1; VDAC1 and VDAC2, voltage-dependent anion channel 1 and 2; and CALU, calumenin isoform c precursor.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: Experimental data-based diagram that rationalizes differentially expressed mitochondrial proteins in a tumor-stroma co-evolution model. ↑: up-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. ↓: down-regulated protein in metastatic 5-8F cells relative to nonmetastatic 6-10B cells. The green frame means the relationship between proteins and tumor metastasis is unclear. The blue frame means the relationship between proteins and tumor metastasis is, at least partially, clear. The blue frame with solid line means protein promotes metastasis and the blue frame with dotted line means protein inhibits metastasis. SOD2, superoxide dismutase [Mn] mitochondrial isoform B; PRDX3, peroxiredoxin 3; PRDX6, peroxiredoxin 6; ECH1, mitochondrial delta(3,5)-delta(2,4)-dienoyl-CoA isomerase; SERPINB5, maspin; COX5A, mitochondrial cytochrome c oxidase subunit 5A; PDIA5, protein disulfide; EIF5A, eukaryotic initiation factor 5A; IDH3B, mitochondrial isocitrate dehydrogenase [NAD] subunit alpha; ETFA, electron transfer flavoprotein; PHB, prohibitin; MRPL12, mitochondrial ribosomal protein L7/L12; PSMC4, 26S protease regulatory subunit 7 isoform 1; VDAC1 and VDAC2, voltage-dependent anion channel 1 and 2; and CALU, calumenin isoform c precursor.
Mentions: Each mitochondrial DEP was rationalized in the tumor-stroma co-evolution model, and an experimental data-based diagram was proposed (Figure9). In our comparative proteomic study of mitochondria between NPC metastatic 5-8F and nonmetastatic 6-10B cells, the first five enriched GO terms of DEPs were closely related to oxidative stress (Table2). For PRDX3, our comparative proteomic data demonstrated the expression of PRDX3 in mitochondria of metastatic 5-8F cells is higher than in mitochondria of nonmetastatic 6-10B cells, it shows that the anti-oxidative stress capability in metastatic 5-8F cells is stronger than nonmetastatic 6-10B cells. This result is consistent with previous conclusion[14] that the enhanced anti-oxidant capabilities in cancer cells defend themselves from excess stromal oxidative damage and favors cell metastasis. Furthermore, among several DEPs (data not showed) selected to carry out siRNA transient transfections and then Transwell Migration assay, only suppression of PRDX3 expression resulted in the significantly increased number of cells migrated through Transwell membrane (P < 0.05). PRDX3, mitochondria thioredoxin-dependent peroxide reductase, is the main way by which cancer cells reduce their levels of H2O2 built up during active respiration[17,18], suppression of this enzyme, a scavenger of peroxide, could increase oxidative stress in cancer cells to up-regulate the redox-signaling, promote the glycolysis, proliferation, and survival, and finally lead increased cells mobility potential[14] (Figure8). To our knowledge, this was the first experimental evidence that PRDX3 was related to NPC metastasis.

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