Identification of melanoma antigens using a Serological Proteome Approach (SERPA).
BACKGROUND
Melanoma is an intractable cancer with a poor prognosis and increasing prevalence worldwide. Specific biomarkers for early diagnosis have yet to be found.


METHODS
Serum samples from melanoma patients and healthy volunteers were utilized for identifying melanoma marker proteins using a serological proteome approach. Specifically, G361 cell protein spots separated by 2-dimensional gel electrophoresis and transferred to a membrane were incubated with patient sera, and positive spots that reacted with more than 5 serum samples were identified using time of flight mass spectrometry.


RESULTS
Only patient sera showed many spots reacted in G361 gels. A total of 13 positive spots were detected and 5 proteins were identified: eukaryotic elongation factor2 (EEF2), enolase1 (ENO1), aldolase A (ALDOA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and heterogeneous nuclear ribonucleoproteins (HNRNP) A2B1. The mRNAs of four proteins (EEF2, ENO1, ALDOA and HNRNPA2B1) were highly expressed in G361 cells compared with melanocytes. EEF2, ENO1 and ALDOA mRNAs were also frequently expressed in other melanoma cell lines.


CONCLUSIONS
The autoantibody-based proteomic approach was effective for investigating melanoma biomarkers. This study might contribute to the development of a diagnostic device for the early detection of cancer.
Suzuki
Ayako
A
Shizuoka Cancer Center Research Institute, Nagaizumi-cho, Sunto-gun, Japan.
Iizuka
Akira
A
Komiyama
Masaru
M
Takikawa
Masako
M
Kume
Akiko
A
Tai
Sachiko
S
Ohshita
Chie
C
Kurusu
Ayumi
A
Nakamura
Youji
Y
Yamamoto
Akifumi
A
Yamazaki
Naoya
N
Yoshikawa
Shusuke
S
Kiyohara
Yoshio
Y
Akiyama
Yasuto
Y
D000951:Antigens, Neoplasm / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D001323:Autoantibodies / Q000097:blood / Q000276:immunology; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D055088:Early Detection of Cancer / Q000379:methods; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000276:immunology* / Q000378:metabolism; D020652:Peptide Elongation Factor 2 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D020543:Proteome / Q000032:analysis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000951:Antigens, Neoplasm; D001323:Autoantibodies; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D020652:Peptide Elongation Factor 2; D020543:Proteome; D012333:RNA, Messenger; D025521:Tumor Suppressor Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005634:Fructose-Bisphosphate Aldolase; C095157:ENO1 protein, human; D010751:Phosphopyruvate Hydratase