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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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Effects of PRDX3 siRNA transient transfection on the PRDX3 expression of metastatic 5-8F cells. Suppression of PRDX3 expression in the levels of mitochondria and whole cells of the metastatic 5-8F cells treated with siRNA relative to without siRNA and control siRNA treatments. The top shows the Western blotting image. The bottom shows the quantitative comparison of the digitized Western blotting image. COXIV, a marker protein of mitochondria, was used as a reference standard protein when mitochondrial protein sample was analyzed with Western blot. ACTIN that is mainly expressed in the cytoplasm was used a reference standard protein when whole-cell protein sample was analyzed with Western blot. Western blot experiments were carried out triply (n = 3).
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Figure 7: Effects of PRDX3 siRNA transient transfection on the PRDX3 expression of metastatic 5-8F cells. Suppression of PRDX3 expression in the levels of mitochondria and whole cells of the metastatic 5-8F cells treated with siRNA relative to without siRNA and control siRNA treatments. The top shows the Western blotting image. The bottom shows the quantitative comparison of the digitized Western blotting image. COXIV, a marker protein of mitochondria, was used as a reference standard protein when mitochondrial protein sample was analyzed with Western blot. ACTIN that is mainly expressed in the cytoplasm was used a reference standard protein when whole-cell protein sample was analyzed with Western blot. Western blot experiments were carried out triply (n = 3).

Mentions: Efficiency of the PRDX3 siRNA (sc-40833) transient transfection of metastatic 5-8F cells was tested with Western Blotting, using a scrambled sequence siRNA (sc-44230) as control. The result data showed PRDX3 expression in mitochondria of metastatic 5-8F cells with PRDX3 siRNA transient transfection was down-regulated (about 65%) compared with metastatic 5-8F cells without PRDX3 siRNA transient transfection (Figure7). It demonstrates that expression of PRDX3 was inhibited in the mitochondria of NPC metastatic 5-8F cells with PRDX3 siRNA treatment.


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 PRDX3 expression of metastatic 5-8F cells. Suppression of PRDX3 expression in the levels of mitochondria and whole cells of the metastatic 5-8F cells treated with siRNA relative to without siRNA and control siRNA treatments. The top shows the Western blotting image. The bottom shows the quantitative comparison of the digitized Western blotting image. COXIV, a marker protein of mitochondria, was used as a reference standard protein when mitochondrial protein sample was analyzed with Western blot. ACTIN that is mainly expressed in the cytoplasm was used a reference standard protein when whole-cell protein sample was analyzed with Western blot. Western blot experiments 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 7: Effects of PRDX3 siRNA transient transfection on the PRDX3 expression of metastatic 5-8F cells. Suppression of PRDX3 expression in the levels of mitochondria and whole cells of the metastatic 5-8F cells treated with siRNA relative to without siRNA and control siRNA treatments. The top shows the Western blotting image. The bottom shows the quantitative comparison of the digitized Western blotting image. COXIV, a marker protein of mitochondria, was used as a reference standard protein when mitochondrial protein sample was analyzed with Western blot. ACTIN that is mainly expressed in the cytoplasm was used a reference standard protein when whole-cell protein sample was analyzed with Western blot. Western blot experiments were carried out triply (n = 3).
Mentions: Efficiency of the PRDX3 siRNA (sc-40833) transient transfection of metastatic 5-8F cells was tested with Western Blotting, using a scrambled sequence siRNA (sc-44230) as control. The result data showed PRDX3 expression in mitochondria of metastatic 5-8F cells with PRDX3 siRNA transient transfection was down-regulated (about 65%) compared with metastatic 5-8F cells without PRDX3 siRNA transient transfection (Figure7). It demonstrates that expression of PRDX3 was inhibited in the mitochondria of NPC metastatic 5-8F cells with PRDX3 siRNA treatment.

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