Differential proteomic analysis of human hepatocellular carcinoma cell line metastasis-associated proteins.
OBJECTIVE
The comparative study of differentially expression of protein profiles of hepatocellular carcinoma cell lines with various metastasic potential and screening key molecules related to hepatocellular carcinoma metastasis and recurrence.


METHODS
Using two-dimensional electrophoresis and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), we analyzed differentially displayed proteomics of human hepatocellular carcinoma cell lines Hep3B, MHCC97L, MHCC97H with different metastasic potential.


RESULTS
Approximate 1,000 protein spots were detected on silver-stained gel by ImageMaster (977+/-113 spots in Hep3B, 1092+/-40 in MHCC97L, and 889+/-14 in MHCC97H). Fifty distinct different protein spots were analyzed with online LC-ESI-MS/MS. Only 26 protein spots had a positive result, including annexin1, S100A4, and so on. In comparison with nonmetastasis Hep3B cell lines, there were 16 proteins overexpressed in MHCC97H and MHCC97L, 10 proteins underexpressed in MHCC97H and MHCC97L. Applying cell immunohistochemistry and RT-PCR, we further validated two interesting and different proteins, annexin1 and S100A4.


CONCLUSIONS
The protein profile of metastatic hepatocellular carcinoma cell lines displayed obvious differences compared with non-metastatic liver cancer cell lines. The results imply that various different proteins may lead to HCC metastasis together.
Cui
Jie-Feng
JF
Liver Cancer Institute, Zhongshan Hospital, Fudan University, 136 Yi Xue Yuan Road, 200032 Shanghai, China.
Liu
Yin-Kun
YK
Pan
Bo-Shen
BS
Song
Hai-Yan
HY
Zhang
Yu
Y
Sun
Rui-Xia
RX
Chen
Jie
J
Feng
Ju-Tao
JT
Tang
Zhao-You
ZY
Yu
Yan-Ling
YL
Shen
Hua-Li
HL
Yang
Peng-Yuan
PY
D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000378:metabolism*; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000032:analysis*; D020543:Proteome; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021241:Spectrometry, Mass, Electrospray Ionization
D009363:Neoplasm Proteins; D020543:Proteome