Generation of novel monoclonal antibodies and their application for detecting ARD1 expression in colorectal cancer
Generation of novel monoclonal antibodies and their application for detecting ARD1 expression in colorectal cancer
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新型单克隆抗体的产生及其在检测结直肠癌中 ARD1 表达中的应用。
DOI：
10.1016/j.canlet.2008.01.028
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发表时间：
2008-06-08
期刊：
影响因子：
9.7
通讯作者：
Shou, Chengchao
中科院分区：
文献类型：
--
作者：
Ren, Tingting;Jiang, Beihai;Shou, Chengchao
Arrest deficient 1（ARD 1）是一种参与酵母细胞周期调控的乙酰转移酶。ARD 1与人N-乙酰转移酶（NATH）相互作用，形成功能性N-末端乙酰转移酶复合物。近年来研究发现它与哺乳动物细胞的增殖和凋亡有关，但其在肿瘤发生发展中的作用尚不清楚。为了评估ARD 1在人结直肠癌中表达的意义，我们产生了一组对ARD 1具有高特异性和灵敏度的单克隆抗体（mAb）。10个克隆均能用于ELISA和Western blot检测，其中克隆10 C12、13 G2和4D 10能与ARD 1在真核细胞中通过免疫沉淀作用相互作用。免疫细胞化学（ICH）和免疫组化（IHC）显示14 D4和10 C12克隆与ARD 1呈强阳性反应。通过使用mAb 14 D4的免疫组织化学分析来评估人结直肠癌和结肠炎组织中的ARD 1表达。50例结直肠癌组织中ARD 1阳性表达41例，而50例正常组织中ARD 1弱阳性表达12例（P < 0.001）。20例结肠炎组织中均未检测到ARD 1表达（P < 0.001）。此外，我们检测的所有六种人结直肠癌细胞系在mRNA和蛋白水平上也是ARD 1阳性的。综上所述，我们产生的抗ARD 1的新型mAb可以为基础和临床研究提供良好的工具，ARD 1可能是结直肠癌的潜在生物标志物。(C)2008爱思唯尔爱尔兰有限公司保留所有权利。
Arrest defective 1 (ARD1) is an acetyltransferase involved in cell cycle control in yeast. ARD1 interacts with human N-acetyltransferase (NATH) to form a functional N-terminal acetyltransferase complex. Recently it had been linked with proliferation and apoptosis in mammalian cells, but its function in cancer development remains unclear. To evaluate significance of ARD1 expression in human colorectal cancer, we generated a panel of monoclonal antibodies (mAbs) with high specificity and sensitivity against ARD1. All of the 10 different clones could be used in ELISA and Western blot, and clone 10C12, 13G2, and 4D10 can interact with ARD1 in eukaryotic cells by immunoprecipitation (IP). Clones of 14D4 and 10C12 were strongly reacted to ARD1 in immunocytochemistry (ICH) and immunohistochemistry (IHC). ARD1 expression was evaluated in human colorectal cancer and colitis tissues by immunohistochemical analysis with mAb 14D4. Forty-one were ARD1-positive in 50 colorectal cancer tissues and only 12 were weak positive in the 50 matched normal tissues (P < 0.001). Moreover, ARD1 expression was not detectable in 20 cases of colitis tissue (P < 0.001). Furthermore, all of the six human colorectal cancer cell lines we examined were also ARD1-positive at mRNA and protein levels. Taken together, the novel mAbs against ARD1 we generated could be good tools for both basic and clinical studies, and ARD1 could be a potential biomarker in colorectal cancer. (C) 2008 Elsevier Ireland Ltd. All rights reserved.