College of Pharmacy, Kyungpook National University, Daegu, Republic of Korea.
School of Pharmacy, Sungkyunkwan University, Suwon, Republic of Korea.
Cancer Genomics Proteomics. 2024 Nov-Dec;21(6):645-652. doi: 10.21873/cgp.20480.
BACKGROUND/AIM: Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer, accounting for approximately 75% of kidney cancers. The objective of this study was to identify novel progression markers for ccRCC based on proteomics, with the goal of stage determination and early diagnosis of kidney cancer patients.
We performed quantitative global proteomics coupled with Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and high-resolution tandem mass spectrometry on kidney-derived cells, including HEK-293, 786-O (primary ccRCC), and Caki-1 (metastatic ccRCC) cells, to investigate the novel progression factors of ccRCC.
In this study, a total of 1,106 proteins were quantified. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted for differentially expressed proteins (DEPs) that were increased in ccRCC cells compared to HEK-293 cells. Ultimately, 99 DEPs including 75 up-regulated and 24 down-regulated proteins, that were significantly altered in both ccRCC cells, were identified. Among DEPs, vimentin was identified as the most significantly changed protein. Its increased expression in ccRCC was verified through immunoblotting in ccRCC cell lines and immunohistochemistry in kidney tumors.
From the global proteomics data detected in ccRCC, we propose 99 DEPs including vimentin as progression factors.
背景/目的:透明细胞肾细胞癌(ccRCC)是最常见的肾癌类型,约占肾癌的 75%。本研究旨在基于蛋白质组学鉴定 ccRCC 的新型进展标志物,以确定肾癌患者的分期和早期诊断。
我们对包括 HEK-293、786-O(原发性 ccRCC)和 Caki-1(转移性 ccRCC)在内的肾源性细胞进行了定量全局蛋白质组学和稳定同位素标记的氨基酸在细胞培养中的应用(SILAC)以及高分辨率串联质谱分析,以研究 ccRCC 的新型进展因子。
在本研究中,共定量了 1106 种蛋白质。对与 HEK-293 细胞相比在 ccRCC 细胞中增加的差异表达蛋白(DEPs)进行了基因本体论和京都基因与基因组百科全书(KEGG)分析。最终,鉴定出 99 种 DEP,包括 75 种上调和 24 种下调蛋白,它们在两种 ccRCC 细胞中均发生明显改变。在 DEP 中,波形蛋白被鉴定为变化最显著的蛋白。通过 ccRCC 细胞系中的免疫印迹和肾脏肿瘤中的免疫组织化学验证了其在 ccRCC 中的高表达。
从 ccRCC 检测到的全局蛋白质组学数据中,我们提出了包括波形蛋白在内的 99 种 DEP 作为进展因子。