Comparative proteomic analysis of human osteosarcoma and SV40-immortalized normal osteoblastic cell lines.
OBJECTIVE
Comparative proteomics provide a powerful approach in screening for alterations in protein levels and post-translational modifications that are associated with tumors. In the present study, we aimed to identify candidate biomarkers to distinguish osteosarcoma (OS) cells from normal osteoblastic cells.


METHODS
We employed 3 OS cell lines (U2OS, IOR/OS9, and SaOS-2), and used the SV40-immortalized normal osteoblastic cell line (hFOB1.19) as the control. The differential protein levels in OS and osteoblastic cells were identified using 2-D gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry analyses. Two proteins of interest, the levels of which were significantly increased in OS cells, were further characterized by Western blot analyses.


RESULTS
Twenty-six proteins were identified, the expression level of which was either significantly increased or decreased in the OS cells as compared to the control cells. The expression level of the activator of 90 kDa shock protein ATPase homolog 1 (AHA1), was enhanced 12.4-, 24.1-, and 23.8-fold in SaOS-2, IOR/OS9, and U2OS cells, respectively, and the level of the stomatin-like protein 2 (SLP-2) was increased by 10.4- and 7.8-fold in IOR/OS9 and U2OS cells, respectively, as compared to normal osteoblastic cells. Those observations were confirmed by Western blot analyses.


CONCLUSIONS
A differential proteomic analysis was successfully used to identify AHA1 and SLP-2 that were significantly overproduced in OS cells as compared to normal osteoblastic cells, suggesting that those proteins among others may be effective biomarker candidates for the identification of OS cells.
Guo
Qian-Chen
QC
Department of Orthopedic Surgery, Sun Yat-sen University, Guangzhou 510080, China.
Shen
Jing-Nan
JN
Jin
Song
S
Wang
Jin
J
Huang
Gang
G
Zhang
Long-Juan
LJ
Huang
Gang
G
Yin
Jun-Qiang
JQ
Zou
Chang-Ye
CY
Li
Ming-Tao
MT
D045744:Cell Line, Tumor / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology*; D012516:Osteosarcoma / Q000235:genetics*; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D013539:Simian virus 40 / Q000235:genetics*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D020543:Proteome