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定量蛋白质表达谱分析 14-3-3 同工型在人肾细胞癌中的表达表明 14-3-3ε参与有限地增加肾细胞增殖。

Quantitative protein expression profiling of 14-3-3 isoforms in human renal carcinoma shows 14-3-3 epsilon is involved in limitedly increasing renal cell proliferation.

机构信息

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, PR China.

出版信息

Electrophoresis. 2009 Dec;30(23):4152-62. doi: 10.1002/elps.200900249.

Abstract

14-3-3 proteins regulate many cellular processes that are implicated in cancer development, and the seven 14-3-3 isoforms have different expression level and isoform-specific roles in different tumors. However, the biological functions of 14-3-3 proteins and their correlations with renal carcinoma have not been investigated so far. In our study, the expression profiles and functional characterization of 14-3-3 proteins were discovered by a sensitive stable isotope labeling with amino acids in cell culture based quantitative proteomics analysis in human renal carcinoma tissues. We found that 14-3-3epsilon was up-regulated with 1.44-fold changes in renal cancerous tissues compared with that in counterpart kidney tissues, and 14-3-3sigma was almost not detected in both tissues due to its DNA highly methylated in our previous reports. The other five isoforms almost have similar expression level in two states of renal tissues. The following RT-PCR, Western blot and immunohistochemistry analysis for specific 14-3-3 isoform expression were all consistent with the quantitative proteomic data. Furthermore, the overexpression of 14-3-3epsilon in vitro can limitedly prompt the abnormal growth of renal tumor cells.

摘要

14-3-3 蛋白调节许多与癌症发展有关的细胞过程,并且这七个 14-3-3 同工型在不同的肿瘤中有不同的表达水平和同工型特异性作用。然而,到目前为止,14-3-3 蛋白的生物学功能及其与肾细胞癌的相关性尚未得到研究。在我们的研究中,通过基于细胞培养的敏感稳定同位素标记与氨基酸定量蛋白质组学分析,在人肾癌细胞组织中发现了 14-3-3 蛋白的表达谱和功能特征。我们发现,与相应的肾组织相比,14-3-3epsilon 在肾癌组织中上调了 1.44 倍,而根据我们之前的报道,由于其 DNA 高度甲基化,14-3-3sigma 在两种组织中几乎无法检测到。其他五种同工型在两种肾组织状态下的表达水平几乎相同。随后的 RT-PCR、Western blot 和免疫组织化学分析特定 14-3-3 同工型的表达情况与定量蛋白质组学数据一致。此外,14-3-3epsilon 的过表达在体外可有限地促使肾肿瘤细胞的异常生长。

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