Medical Oncology Department, The First Affiliated Hospital, College of Medicine, Xi'an Jiaotong University, Xi'an 710061, P.R. China.
Int J Oncol. 2013 Jun;42(6):2078-86. doi: 10.3892/ijo.2013.1889. Epub 2013 Apr 10.
Identification of new therapeutic and prognostic biomarkers for clear cell renal cell carcinoma (ccRCC) is urgently required since most patients are in advanced stages of ccRCC at initial diagnosis and the recurrence rate is high. Differentially expressed proteins between the ccRCC cell line RLC-310 and the normal renal cell line HK-2 were identified by a comparative proteomic approach based on a protein fractionation two-dimensional (PF-2D) liquid-phase fractionation system and capillary liquid chromatography electrospray ionization mass spectrometry/mass spectrometry (LC-ESI-MS/MS). Differentially expressed proteins (n=196) were identified. Of the 13 differentially expressed proteins newly discovered in ccRCC, angiomotin (Amot) was the focus of this study since its role in ccRCC progression has been obscure. The overexpression of Amot in ccRCC tissues was confirmed by comparing Amot expression in 18 primary ccRCC tissues and adjacent normal renal tissues (ANRT) using western blot analysis. Quantitative RT-PCR using 127 ccRCC tissues revealed that high levels of Amot transcripts were associated with poor differentiation, venous invasion and decreased survival (p<0.0001, <0.05 and <0.05). Multivariate analysis indicated that Amot transcript was an independent prognostic factor for the survival of ccRCC patients (p<0.05). These data suggest that Amot may serve as a novel prognostic factor of ccRCC.
由于大多数患者在初始诊断时已经处于 ccRCC 的晚期，且复发率较高，因此迫切需要鉴定新的治疗和预后生物标志物用于透明细胞肾细胞癌（ccRCC）。本研究采用基于蛋白分级二维（PF-2D）液相分级系统和毛细管液相色谱电喷雾电离质谱/质谱（LC-ESI-MS/MS）的比较蛋白质组学方法鉴定了 ccRCC 细胞系 RLC-310 和正常肾细胞系 HK-2 之间的差异表达蛋白。鉴定出差异表达蛋白（n=196）。在新发现的 13 个在 ccRCC 中差异表达的蛋白中，血管生成素（Amot）是本研究的重点，因为其在 ccRCC 进展中的作用尚不清楚。通过 Western blot 分析比较 18 个原发性 ccRCC 组织和相邻正常肾组织（ANRT）中的 Amot 表达，证实了 Amot 在 ccRCC 组织中的过表达。对 127 个 ccRCC 组织进行的定量 RT-PCR 显示，Amot 转录本水平高与分化不良、静脉浸润和生存时间缩短相关（p<0.0001、<0.05 和 <0.05）。多变量分析表明，Amot 转录本是 ccRCC 患者生存的独立预后因素（p<0.05）。这些数据表明 Amot 可能是 ccRCC 的一个新的预后因素。
