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Tumor Biology

, Volume 36, Issue 4, pp 2973–2982 | Cite as

MDA-9 and GRP78 as potential diagnostic biomarkers for early detection of melanoma metastasis

Research Article

Abstract

Metastatic melanoma, the primary cause of skin cancer-related death, warrants new diagnostic and therapeutic approaches that target the regulatory machinery at molecular level. The heterogeneity and complexity of melanoma result in the difficulty to find biomarkers and targets for early detection and treatment. Here, we investigated metastasis-associated proteins by comparing the proteomic profiles of primary cutaneous melanomas to their matched lymph node metastases, which minimizes heterogeneity among samples from different patients. Results of two-dimensional gel electrophoresis (2-DE) followed by proteomic analysis revealed eight differentially expressed proteins. Among them, seven proteins (α-enolase, cofilin-1, LDH, m-β-actin, Nm23, GRP78, and MDA-9) showed increased and one (annexin A2) showed decreased expression in metastatic lymph node tissues than in primary melanomas. MDA-9 and GRP78 were the most highly expressed proteins in lymph node metastases, which was validated by immunohistochemical staining. Moreover, exosomes from serum samples of metastatic melanoma patients contained higher levels of MDA-9 and GRP78 than those of patients without metastases, indicating the potential of MDA-9 and GRP78 to be biomarkers for early detection of metastasis. Further, small interfering RNA (siRNA)-mediated knockdown confirmed a functional role for MDA-9 and GRP78 to promote cell invasion in the A375 cells. Finally, we showed that GRP78 co-localized with MDA-9 in 293T cells. Taken together, our findings support MDA-9, co-expressed with GRP78, as a melanoma protein associated with lymph node metastasis. Investigating how MDA-9 and GRP78 interact to contribute to melanoma metastasis and disease progression could reveal new potential avenues of targeted therapy and/or useful biomarkers for diagnosis and prognosis.

Keywords

Melanoma MDA-9 GRP78 Exosomes Two-dimensional gel electrophoresis 

Notes

Acknowledgments

The study was supported, in whole or in part, by the National Natural Science Foundation of China no. 81272380 (to B.S.), no. 81272386 (to M.G.), and the Research Grants of Shenzhen Science and Technology project ZYA201106080030A and KQCX20120803145850990. The authors would like to thank Shenzhen Biomedical Research Support Platform for their excellent technical assistance.

Conflict of interest

None

Supplementary material

13277_2014_2930_MOESM1_ESM.doc (30 kb)
ESM 1 (DOC 30 kb)

References

  1. 1.
    Diepgen TL, Mahler V. The epidemiology of skin cancer. Br J Dermatol. 2002;146 Suppl 61:1–6.CrossRefPubMedGoogle Scholar
  2. 2.
    Little EG, Eide MJ. Update on the current state of melanoma incidence. Dermatol Clin. 2012;30:355–61.CrossRefPubMedGoogle Scholar
  3. 3.
    Balch CM, Sober AJ, Soong SJ, Gershenwald JE, Committee AMS. The new melanoma staging system. Semin Cutan Med Surg. 2003;22:42–54.CrossRefPubMedGoogle Scholar
  4. 4.
    Berger MF, Levin JZ, Vijayendran K, Sivachenko A, Adiconis X, Maguire J, et al. Integrative analysis of the melanoma transcriptome. Genome Res. 2010;20:413–27.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Pleasance ED, Cheetham RK, Stephens PJ, McBride DJ, Humphray SJ, Greenman CD, et al. A comprehensive catalogue of somatic mutations from a human cancer genome. Nat. 2010;463:191–6.CrossRefGoogle Scholar
  6. 6.
    Valenti R, Huber V, Filipazzi P, Pilla L, Sovena G, Villa A, et al. Human tumor-released microvesicles promote the differentiation of myeloid cells with transforming growth factor-beta-mediated suppressive activity on t lymphocytes. Cancer Res. 2006;66:9290–8.CrossRefPubMedGoogle Scholar
  7. 7.
    Andre F, Schartz NE, Movassagh M, Flament C, Pautier P, Morice P, et al. Malignant effusions and immunogenic tumour-derived exosomes. Lancet. 2002;360:295–305.CrossRefPubMedGoogle Scholar
  8. 8.
    Vlassov AV, Magdaleno S, Setterquist R, Conrad R. Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim Biophys Acta. 1820;2012:940–8.Google Scholar
  9. 9.
    Xiao D, Ohlendorf J, Chen Y, Taylor DD, Rai SN, Waigel S, et al. Identifying mRNA, microRNA and protein profiles of melanoma exosomes. PLoS One. 2012;7:e46874.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Nicholas J. A new diagnostic tool with the potential to predict tumor metastasis. J Natl Cancer Inst. 2013;105:371–2.CrossRefPubMedGoogle Scholar
  11. 11.
    Skog J, Wurdinger T, van Rijn S, Meijer DH, Gainche L, Sena-Esteves M, et al. Glioblastoma microvesicles transport rna and proteins that promote tumour growth and provide diagnostic biomarkers. Nat Cell Biol. 2008;10:1470–6.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Skokos D, Botros HG, Demeure C, Morin J, Peronet R, Birkenmeier G, et al. Mast cell-derived exosomes induce phenotypic and functional maturation of dendritic cells and elicit specific immune responses in vivo. J Immunol. 2003;170:3037–45.CrossRefPubMedGoogle Scholar
  13. 13.
    Escola JM, Kleijmeer MJ, Stoorvogel W, Griffith JM, Yoshie O, Geuze HJ. Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. J Biol Chem. 1998;273:20121–7.CrossRefPubMedGoogle Scholar
  14. 14.
    Wubbolts R, Leckie RS, Veenhuizen PT, Schwarzmann G, Mobius W, Hoernschemeyer J, et al. Proteomic and biochemical analyses of human B cell-derived exosomes. Potential implications for their function and multivesicular body formation. J Biol Chem. 2003;278:10963–72.CrossRefPubMedGoogle Scholar
  15. 15.
    Nakayama T, Taback B, Turner R, Morton DL, Hoon DS. Molecular clonality of in-transit melanoma metastasis. Am J Pathol. 2001;158:1371–8.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Daigo Y, Chin SF, Gorringe KL, Bobrow LG, Ponder BA, Pharoah PD, et al. Degenerate oligonucleotide primed-polymerase chain reaction-based array comparative genomic hybridization for extensive amplicon profiling of breast cancers: a new approach for the molecular analysis of paraffin-embedded cancer tissue. Am J Pathol. 2001;158:1623–31.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Peinado H, Aleckovic M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through met. Nat Med. 2012;18:883–91.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Su B, Bu Y, Engelberg D, Gelman IH. Ssecks/gravin/akap12 inhibits cancer cell invasiveness and chemotaxis by suppressing a protein kinase C- Raf/MEK/ERK pathway. J Biol Chem. 2010;285:4578–86.CrossRefPubMedGoogle Scholar
  19. 19.
    Hwangbo C, Kim J, Lee JJ, Lee JH. Activation of the integrin effector kinase focal adhesion kinase in cancer cells is regulated by crosstalk between protein kinase calpha and the pdz adapter protein mda-9/syntenin. Cancer Res. 2010;70:1645–55.CrossRefPubMedGoogle Scholar
  20. 20.
    Boukerche H, Su ZZ, Prevot C, Sarkar D, Fisher PB. Mda-9/syntenin promotes metastasis in human melanoma cells by activating c-src. Proc Natl Acad Sci U S A. 2008;105:15914–9.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Papalas JA, Vollmer RT, Gonzalez-Gronow M, Pizzo SV, Burchette J, Youens KE, et al. Patterns of grp78 and mtj1 expression in primary cutaneous malignant melanoma. Mod Pathol: Off J U S Can Acad Pathol Inc. 2010;23:134–43.CrossRefGoogle Scholar
  22. 22.
    Zhuang L, Scolyer RA, Lee CS, McCarthy SW, Cooper WA, Zhang XD, et al. Expression of glucose-regulated stress protein grp78 is related to progression of melanoma. Histopathol. 2009;54:462–70.CrossRefGoogle Scholar
  23. 23.
    Selim MA, Burchette JL, Bowers EV, de Ridder GG, Mo L, Pizzo SV, et al. Changes in oligosaccharide chains of autoantibodies to grp78 expressed during progression of malignant melanoma stimulate melanoma cell growth and survival. Melanoma Res. 2011;21:323–34.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Vandenbroeck K, Alloza I, Brehmer D, Billiau A, Proost P, McFerran N, et al. The conserved helix c region in the superfamily of interferon-gamma /interleukin-10-related cytokines corresponds to a high-affinity binding site for the hsp70 chaperone dnak. J Biol Chem. 2002;277:25668–76.CrossRefPubMedGoogle Scholar
  25. 25.
    Huang SK, Darfler MM, Nicholl MB, You J, Bemis KG, Tegeler TJ, et al. Lc/ms-based quantitative proteomic analysis of paraffin-embedded archival melanomas reveals potential proteomic biomarkers associated with metastasis. PLoS One. 2009;4:e4430.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Hardesty WM, Kelley MC, Mi D, Low RL, Caprioli RM. Protein signatures for survival and recurrence in metastatic melanoma. J Proteome. 2011;74:1002–14.CrossRefGoogle Scholar
  27. 27.
    Wang P, Bouwman FG, Mariman EC. Generally detected proteins in comparative proteomics—a matter of cellular stress response? Proteome. 2009;9:2955–66.CrossRefGoogle Scholar
  28. 28.
    Boukerche H, Su ZZ, Emdad L, Baril P, Balme B, Thomas L, et al. Mda-9/syntenin: a positive regulator of melanoma metastasis. Cancer Res. 2005;65:10901–11.CrossRefPubMedGoogle Scholar
  29. 29.
    Gangemi R, Mirisola V, Barisione G, Fabbi M, Brizzolara A, Lanza F, et al. Mda-9/syntenin is expressed in uveal melanoma and correlates with metastatic progression. PLoS One. 2012;7:e29989.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Fernandez-Larrea J, Merlos-Suarez A, Urena JM, Baselga J, Arribas J. A role for a pdz protein in the early secretory pathway for the targeting of protgf-alpha to the cell surface. Mol Cell. 1999;3:423–33.CrossRefPubMedGoogle Scholar
  31. 31.
    Misra UK, Gonzalez-Gronow M, Gawdi G, Wang F, Pizzo SV. A novel receptor function for the heat shock protein grp78: silencing of grp78 gene expression attenuates alpha2m*-induced signalling. Cell Signal. 2004;16:929–38.CrossRefPubMedGoogle Scholar
  32. 32.
    Misra UK, Deedwania R, Pizzo SV. Binding of activated alpha2-macroglobulin to its cell surface receptor grp78 in 1-ln prostate cancer cells regulates pak-2-dependent activation of limk. J Biol Chem. 2005;280:26278–86.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Ming Guan
    • 1
  • Xiaofan Chen
    • 2
    • 3
  • Yingyu Ma
    • 4
  • Lihua Tang
    • 5
    • 6
  • Lei Guan
    • 7
  • Xuefeng Ren
    • 8
  • Bo Yu
    • 3
    • 5
    • 9
  • Wei Zhang
    • 2
    • 3
    • 9
  • Bing Su
    • 2
    • 3
    • 8
    • 9
  1. 1.Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical CollegeFudan UniversityShanghaiChina
  2. 2.Biomedical Research InstituteShenzhen Peking University - The Hong Kong University of Science and Technology Medical CenterGuangdongChina
  3. 3.Shenzhen Key Laboratory for Translational Medicine of Dermatology, Biomedical Research InstituteShenzhen Peking University - The Hong Kong University of Science and Technology Medical CenterGuangdongChina
  4. 4.Department of Pharmacology and TherapeuticsRoswell Park Cancer InstituteBuffaloUSA
  5. 5.Department of DermatologyPeking University Shenzhen HospitalShenzhenChina
  6. 6.Department of Dermatology303 Hospital of People’s Liberation Army of ChinaNanningChina
  7. 7.Skin Research Center of Guangzhou LandproofGuangzhouChina
  8. 8.Department of Epidemiology and Environmental Health, School of Public Health and Health ProfessionsThe State University of New YorkBuffaloUSA
  9. 9.Shenzhen Key Discipline of DermatologyPeking University Shenzhen HospitalShenzhenChina

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