Guo Qian-Chen, Shen Jing-Nan, Jin Song, Wang Jin, Huang Gang, Zhang Long-Juan, Huang Gang, Yin Jun-Qiang, Zou Chang-Ye, Li Ming-Tao
Department of Orthopedic Surgery, Sun Yat-sen University, Guangzhou 510080, China.
Acta Pharmacol Sin. 2007 Jun;28(6):850-8. doi: 10.1111/j.1745-7254.2007.00603.x.
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.
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.
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.
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.
比较蛋白质组学为筛选与肿瘤相关的蛋白质水平变化和翻译后修饰提供了一种强大的方法。在本研究中，我们旨在鉴定候选生物标志物以区分骨肉瘤（OS）细胞与正常成骨细胞。
我们采用了3种OS细胞系（U2OS、IOR/OS9和SaOS-2），并使用SV40永生化正常成骨细胞系（hFOB1.19）作为对照。通过二维凝胶电泳，随后进行基质辅助激光解吸/电离飞行时间质谱分析，鉴定OS细胞和成骨细胞中的差异蛋白质水平。对两种在OS细胞中水平显著升高的感兴趣蛋白质，通过蛋白质印迹分析进一步进行表征。
鉴定出26种蛋白质，与对照细胞相比，其在OS细胞中的表达水平显著升高或降低。90 kDa热休克蛋白ATP酶同源物1（AHA1）的激活剂在SaOS-2、IOR/OS9和U2OS细胞中的表达水平分别提高了12.4倍、24.1倍和23.8倍，与正常成骨细胞相比，类stomatin蛋白2（SLP-2）在IOR/OS9和U2OS细胞中的水平分别提高了10.4倍和7.8倍。这些观察结果通过蛋白质印迹分析得到证实。
差异蛋白质组学分析成功用于鉴定与正常成骨细胞相比在OS细胞中显著过量产生的AHA1和SLP-2，表明这些蛋白质以及其他蛋白质可能是鉴定OS细胞的有效候选生物标志物。
