Identification of metastasis candidate proteins among HCC cell lines by comparative proteome and biological function analysis of S100A4 in metastasis in vitro.
Widespread metastasis of hepatocellular carcinoma (HCC) was a complex cascade of events, which is still beyond full appreciation. Screening key proteins, which play a critical role in metastasis, using high-throughput proteomics approach help discover valuable biomarkers and elucidate the mechanism of metastasis. This study was to find out some metastasis candidate proteins among HCC cell lines with various metastatic potential by comparative proteomics, and then further validate the biological function of these proteins in metastasis in vitro. The protein profiles of metastatic HCC cell lines (MHCC97H and MHCC97L) displayed obvious differences compared with nonmetastatic ones (Hep3B). Twenty-six metastasis candidate proteins, which were identified by on-line LC-ESI-MS/MS, such as S100 calcium-binding protein A4 (S100A4), annexin 1, etc., might have much application in diagnostic procedures and prognosis evaluation. S100A4, as a leading different metastasis candidate protein, which overexpressed only in the metastatic cells, was selected for further investigation. A series of assays related to invasion and metastasis in vitro, including cell motility, invasion, and matrix metalloproteinases (MMPs) secretion, were performed in MHCC97H/antisense recombinant plasmid to S100A4 (pcDNA3.1(+) AS S100A4) and the mock controls. All the data in the present study suggested that S100A4 might contribute to HCC invasion and metastasis through two paths of matrix metalloproteinase (MMP9) secretion regulation and strengthened motility and invasion properties.
Cui
Jie Feng
JF
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, P. R. China.
Liu
Yin Kun
YK
Zhang
Li Jun
LJ
Shen
Hua Li
HL
Song
Hai Yan
HY
Dai
Zhi
Z
Yu
Yan Ling
YL
Zhang
Yu
Y
Sun
Rui Xia
RX
Chen
Jie
J
Tang
Zhao You
ZY
Yang
Peng Yuan
PY
D017209:Apoptosis; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D003470:Culture Media / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007120:Immunochemistry; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D013058:Mass Spectrometry / Q000379:methods; D020782:Matrix Metalloproteinases / Q000378:metabolism; D009361:Neoplasm Invasiveness; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000378:metabolism*; D014162:Transfection
D003470:Culture Media; D020319:Oligodeoxyribonucleotides, Antisense; D020543:Proteome; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C077591:S100A4 protein, human; D004262:DNA Restriction Enzymes; D020782:Matrix Metalloproteinases