https://scholars.lib.ntu.edu.tw/handle/123456789/454719
標題: | Antibody-assisted target identification reveals afatinib, an EGFR covalent inhibitor, down-regulating ribonucleotide reductase | 作者: | Yu C.-H. Chou C.-C. Tu H.-F. Huang W.-C. Ho Y.-Y. Khoo K.-H. MING-SHYUE LEE GEEN-DONG CHANG |
公開日期: | 2018 | 出版社: | Impact Journals LLC | 卷: | 9 | 期: | 30 | 起(迄)頁: | 21512-21529 | 來源出版物: | Oncotarget | 摘要: | Afatinib, used for the first-line treatment of non-small-cell lung carcinoma (NSCLC) patients with distinct epidermal growth factor receptor (EGFR) mutations, inactivates EGFR by mimicking ATP structure and forming a covalent adduct with EGFR. We developed a method to unravel potential targets of afatinib in NSCLC cells through immunoprecipitation of afatinib-labeling proteins with anti-afatinib antiserum and mass spectrometry analysis. Ribonucleotide reductase (RNR) is one of target proteins of afatinib revealed by this method. Treatment of afatinib at 10-100 nM potently inhibited intracellular RNR activity in an in vitro assay using permeabilized PC-9 cells (formerly known as PC-14). PC-9 cells treated with 10 μM afatinib displayed elevated markers of DNA damage. Long-term treatment of therapeutic concentrations of afatinib in PC-9 cells caused significant decrease in protein levels of RNR subunit M2 at 1-10 nM and RNR subunit M1 at 100 nM. EGFR-null Chinese hamster ovary (CHO) cells treated with afatinib also showed similar effects. Afatinib repressed the upregulation of RNR subunit M2 induced by gemcitabine. Covalent modification with afatinib resulting in inhibition and protein downregulation of RNR underscores the therapeutic and off-target effects of afatinib. Afatinib may serve as a lead compound of chemotherapeutic drugs targeting RNR. This method can be widely used in the identification of potential targets of other covalent drugs. ? Yu et al. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85045842622&doi=10.18632%2foncotarget.25177&partnerID=40&md5=2ca9a99f8317e20f74dd53fcb35c16ef https://scholars.lib.ntu.edu.tw/handle/123456789/454719 |
ISSN: | 1949-2553 | DOI: | 10.18632/oncotarget.25177 | SDG/關鍵字: | afatinib; DNA; gemcitabine; ribonucleotide reductase; animal cell; animal experiment; animal model; animal tissue; Article; CHO cell line; controlled study; DNA damage; down regulation; drug targeting; female; in vitro study; infant; male; mouse; non small cell lung cancer; nonhuman; PC-9 cell line; protein analysis; upregulation |
顯示於: | 生物化學暨分子生物學科研究所 |
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