Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer
About 80% of lung cancer patients are diagnosed with non–small cell lung cancer (NSCLC). EGFR mutation and overexpression are common in NSCLC, thus making EGFR signaling a key target for therapy. While EGFR kinase inhibitors (EGFR–TKIs) are widely used and efficacious in treatment, increases in resi...
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| Format: | Article |
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MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/19/14879 |
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| author | Ya-Yu Yang Sheng-Chieh Lin Jong-Ding Lay Chun-Yu Cho Te-Hsuan Jang Hsiu-Ying Ku Chih-Jung Yao Shuang-En Chuang |
| author_facet | Ya-Yu Yang Sheng-Chieh Lin Jong-Ding Lay Chun-Yu Cho Te-Hsuan Jang Hsiu-Ying Ku Chih-Jung Yao Shuang-En Chuang |
| author_sort | Ya-Yu Yang |
| collection | DOAJ |
| description | About 80% of lung cancer patients are diagnosed with non–small cell lung cancer (NSCLC). EGFR mutation and overexpression are common in NSCLC, thus making EGFR signaling a key target for therapy. While EGFR kinase inhibitors (EGFR–TKIs) are widely used and efficacious in treatment, increases in resistance and tumor recurrence with alternative survival pathway activation, such as that of AXL and MET, occur frequently. AXL is one of the EMT (epithelial–mesenchymal transition) signature genes, and EMT morphological changes are also responsible for EGFR–TKI resistance. MIG6 is a negative regulator of ERBB signaling and has been reported to be positively correlated with EGFR–TKI resistance, and downregulation of MIG6 by miR–200 enhances EMT transition. While MIG6 and AXL are both correlated with EMT and EGFR signaling pathways, how AXL, MIG6 and EGFR interplay in lung cancer remains elusive. Correlations between AXL and MIG6 expression were analyzed using Oncomine or the CCLE. A luciferase reporter assay was used for determining MIG6 promoter activity. Ectopic overexpression, RNA interference, Western blot analysis, qRT–PCR, a proximity ligation assay and a coimmunoprecipitation assay were performed to analyze the effects of certain gene expressions on protein–protein interaction and to explore the underlying mechanisms. An in vitro kinase assay and LC–MS/MS were utilized to determine the phosphorylation sites of AXL. In this study, we demonstrate that MIG6 is a novel substrate of AXL and is stabilized upon phosphorylation at Y310 and Y394/395 by AXL. This study reveals a connection between MIG6 and AXL in lung cancer. AXL phosphorylates and stabilizes MIG6 protein, and in this way EGFR signaling may be modulated. This study may provide new insights into the EGFR regulatory network and may help to advance cancer treatment. |
| first_indexed | 2024-03-10T21:43:01Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T21:43:01Z |
| publishDate | 2023-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b2190d3f225241f7b1468536fb899ca52023-11-19T14:31:59ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-10-0124191487910.3390/ijms241914879Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung CancerYa-Yu Yang0Sheng-Chieh Lin1Jong-Ding Lay2Chun-Yu Cho3Te-Hsuan Jang4Hsiu-Ying Ku5Chih-Jung Yao6Shuang-En Chuang7National Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanNational Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanDepartment of Nursing, National Taichung University of Science and Technology, Taichung 40343, TaiwanNational Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanNational Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanNational Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanDepartment of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, TaiwanNational Institute of Cancer Research, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli 35053, TaiwanAbout 80% of lung cancer patients are diagnosed with non–small cell lung cancer (NSCLC). EGFR mutation and overexpression are common in NSCLC, thus making EGFR signaling a key target for therapy. While EGFR kinase inhibitors (EGFR–TKIs) are widely used and efficacious in treatment, increases in resistance and tumor recurrence with alternative survival pathway activation, such as that of AXL and MET, occur frequently. AXL is one of the EMT (epithelial–mesenchymal transition) signature genes, and EMT morphological changes are also responsible for EGFR–TKI resistance. MIG6 is a negative regulator of ERBB signaling and has been reported to be positively correlated with EGFR–TKI resistance, and downregulation of MIG6 by miR–200 enhances EMT transition. While MIG6 and AXL are both correlated with EMT and EGFR signaling pathways, how AXL, MIG6 and EGFR interplay in lung cancer remains elusive. Correlations between AXL and MIG6 expression were analyzed using Oncomine or the CCLE. A luciferase reporter assay was used for determining MIG6 promoter activity. Ectopic overexpression, RNA interference, Western blot analysis, qRT–PCR, a proximity ligation assay and a coimmunoprecipitation assay were performed to analyze the effects of certain gene expressions on protein–protein interaction and to explore the underlying mechanisms. An in vitro kinase assay and LC–MS/MS were utilized to determine the phosphorylation sites of AXL. In this study, we demonstrate that MIG6 is a novel substrate of AXL and is stabilized upon phosphorylation at Y310 and Y394/395 by AXL. This study reveals a connection between MIG6 and AXL in lung cancer. AXL phosphorylates and stabilizes MIG6 protein, and in this way EGFR signaling may be modulated. This study may provide new insights into the EGFR regulatory network and may help to advance cancer treatment.https://www.mdpi.com/1422-0067/24/19/14879AXLMIG6ERRFI1EGFRNSCLC |
| spellingShingle | Ya-Yu Yang Sheng-Chieh Lin Jong-Ding Lay Chun-Yu Cho Te-Hsuan Jang Hsiu-Ying Ku Chih-Jung Yao Shuang-En Chuang Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer International Journal of Molecular Sciences AXL MIG6 ERRFI1 EGFR NSCLC |
| title | Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer |
| title_full | Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer |
| title_fullStr | Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer |
| title_full_unstemmed | Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer |
| title_short | Intervention of AXL in EGFR Signaling via Phosphorylation and Stabilization of MIG6 in Non-Small Cell Lung Cancer |
| title_sort | intervention of axl in egfr signaling via phosphorylation and stabilization of mig6 in non small cell lung cancer |
| topic | AXL MIG6 ERRFI1 EGFR NSCLC |
| url | https://www.mdpi.com/1422-0067/24/19/14879 |
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