https://scholars.lib.ntu.edu.tw/handle/123456789/507231
標題: | Small molecule T315 promotes casitas b-lineage lymphoma dependent degradation of epidermal growth factor receptor via Y1045 autophosphorylation | 作者: | Huang K.-Y. Kao S.-H. Wang W.-L. Chen C.-Y. Hsiao T.-H. Salunke S.B. Chen J.J.W. KANG-YI SU Yang S.-C. Hong T.-M. Chen C.-S. PAN-CHYR YANG |
公開日期: | 2016 | 卷: | 193 | 期: | 7 | 起(迄)頁: | 753-766 | 來源出版物: | American Journal of Respiratory and Critical Care Medicine | 摘要: | Rationale: Despite the fact that tyrosine kinase inhibitors (TKIs) have been found effective in treating patients harboring activating mutations of epidermal growth factor receptor (EGFR), an acquired secondary mutation, T790M, which lowers the affinity to TKIs, can lead to EGFR TKI resistance after this standard treatment. Objectives: To evaluate the effect of small molecule T315 on EGFR degradation and its therapeutic efficacy in vitro and in vivo. Methods: Lung adenocarcinoma cells were treated with T315, and cell proliferation and apoptotic proportion were determined by the CellTiter 96 AQueous MTS (3-[4,5-dimethylthiazol-2-yl]-5- [3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, inner salt) assay and flow cytometry. The effects of T315 on EGFR mRNA and protein levels, autophosphorylation, ubiquitination, and degradation were evaluated by real-time polymerase chain reaction and Western blot, respectively. Direct targeting of T315 to EGFR was confirmed bythe in vitro kinase assayandmass spectrometry. Finally, the preclinical effect of T315 was validated in the murine xenograft model in combination with a second-generation TKI, afatinib. Measurements and Main Results: We identified T315 as a novel, potent small molecule for suppressing cancer cell proliferation in vitro and in vivo. The therapeutic effect was verified after T315 was combined with a second-generation TKI, afatinib, compared with a single drug administration. We found a new mechanism of action, in that T315 appears to directly bind EGFR and triggers EGFR-Y1045 autophosphorylation, whereby its degradation is triggered through the ubiquitin-proteasome pathway. Conclusions: Our evidence suggests that T315 is a novel class of anticancerdrugthatisabletoinhibitthegrowthofEGFR-TKI resistant lung adenocarcinoma cells by inducing the degradation of EGFR. ? 2016 by the American Thoracic Society. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/507231 | ISSN: | 1073-449X | DOI: | 10.1164/rccm.201502-0250OC | SDG/關鍵字: | afatinib; antineoplastic agent; epidermal growth factor receptor; proteasome; protein tyrosine kinase inhibitor; t 315; ubiquitin; unclassified drug; afatinib; antineoplastic agent; Cbl protein; Cblb protein, mouse; drug combination; EGFR protein, human; epidermal growth factor receptor; protein kinase inhibitor; quinazoline derivative; signal transducing adaptor protein; animal model; antiproliferative activity; Article; autophosphorylation; binding affinity; cancer resistance; cell survival; controlled study; down regulation; drug efficacy; drug mechanism; human; human cell; IC50; in vitro study; in vivo study; lung adenocarcinoma; mouse; nonhuman; priority journal; protein degradation; protein expression; protein interaction; ubiquitination; adenocarcinoma; animal; antagonists and inhibitors; cell proliferation; drug combination; drug effects; drug resistance; drug screening; enzyme assay; genetics; Lung Neoplasms; mass spectrometry; mutation; real time polymerase chain reaction; tumor cell culture; Western blotting; Adaptor Proteins, Signal Transducing; Adenocarcinoma; Animals; Antineoplastic Agents; Blotting, Western; Cell Proliferation; Drug Combinations; Drug Resistance, Neoplasm; Enzyme Assays; Humans; Lung Neoplasms; Mass Spectrometry; Mice; Mutation; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-cbl; Quinazolines; Real-Time Polymerase Chain Reaction; Receptor, Epidermal Growth Factor; Tumor Cells, Cultured; Xenograft Model Antitumor Assays |
顯示於: | 醫學檢驗暨生物技術學系 |
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