[Screening hepatocellular carcinoma autoantibodies by serological proteome analysis].
OBJECTIVE
To screen hepatocellular carcinoma (HCC) autoantibodies as diagnostic biomarkers or therapy targets by serologic proteome analysis (SERPA).


METHODS
Total proteins extracted from human HCC cell line HCCLM3 were separated by two-dimensional electrophoresis (2-DE) and then transferred onto PVDF membranes, which were subsequently incubated with sera from HCC, hepatitis B virus (HBV) infected patients or healthy volunteers. All immuno-reactive protein spots on blot films were matched to those on 2-DE gel maps by image analysis and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS/MS).


RESULTS
2-DE gel maps of HCCLM3 and corresponding blot films of good quality and reproducibility were established. The number of spots on HCCLM3 2-DE reference gel totaled 603 and those on HCC, HBV and healthy sera blotted films were 70.75+/-24.25, 68.5+/-23.44 and 41.38+/-15.05, respectively. Blot films of HCC and HBV groups had more spots than those of the healthy group (P < 0.05) while no significance was found between films of HCC and HBV groups. By identification, those HCC autoantibodies could be classified as nuclear proteins, cytoskeleton proteins, heat shock proteins and metabolic enzymes.


CONCLUSIONS
Serological proteome analysis is a high throughput technique for screening tumor autoantibodies. Those newly identified HCC associated tumor antigens and corresponding autoantibodies can be used in the early diagnosis or immuno-therapy of HCC.
Feng
Ju-tao
JT
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China. fengjt@hotmail.com
Liu
Yin-kun
YK
Dai
Zhi
Z
Zhou
Hai-jun
HJ
Song
Hai-yan
HY
Qin
Lun-xiu
LX
Jin
Hong
H
Lu
Hao-jie
HJ
Tang
Zhao-you
ZY
D000912:Antibodies, Neoplasm / Q000032:analysis*; D001323:Autoantibodies / Q000032:analysis*; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008113:Liver Neoplasms / Q000276:immunology*; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014407:Tumor Cells, Cultured
D000912:Antibodies, Neoplasm; D001323:Autoantibodies