EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation
Aims: The aim of this study was to reveal the specific molecular mechanisms by which DENND1A accepts EGF signaling and activates Rab35 in gastric cancer.Methods: The expression of proteins related to DENND1A was examined by western blot analysis. Activation of Rab35 was assessed by GST-pulldown. The...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2018-11-01
|
| Series: | Frontiers in Pharmacology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fphar.2018.01343/full |
| _version_ | 1819261590556377088 |
|---|---|
| author | Bixing Ye Biao Duan Wenjie Deng Yueyuan Wang Yan Chen Jie Cui Jie Cui Shixiu Sun Yujie Zhang Yujie Zhang Jun Du Jun Du Luo Gu Luo Gu Luo Gu Lin Lin Yurong Tang |
| author_facet | Bixing Ye Biao Duan Wenjie Deng Yueyuan Wang Yan Chen Jie Cui Jie Cui Shixiu Sun Yujie Zhang Yujie Zhang Jun Du Jun Du Luo Gu Luo Gu Luo Gu Lin Lin Yurong Tang |
| author_sort | Bixing Ye |
| collection | DOAJ |
| description | Aims: The aim of this study was to reveal the specific molecular mechanisms by which DENND1A accepts EGF signaling and activates Rab35 in gastric cancer.Methods: The expression of proteins related to DENND1A was examined by western blot analysis. Activation of Rab35 was assessed by GST-pulldown. The interaction of DENND1A and Grb2 was assessed by GST-pulldown and co-immunoprecipitation assays. The relationship between DENND1A and cell migration and invasion was detected using wound healing and transwell by gene overexpression and RNA interference.Results: EGF stimulation significantly promoted cell migration, whereas transfection with siRab35 partially inhibited EGF-promoted cell migration. DENND1A is also involved in these processes and active Rab35. Moreover, DENND1A binds to the N-terminal and C-terminal SH3 domains of Grb2 through PRD. Of special interest is the observation that EGFR can recruit Grb2-DENND1A complex under EGF stimulation. Further results reveal that the higher the expression of DENND1A, the shorter progression-free survival of gastric cancer patients.Conclusion: In summary, we confirmed that EGF-Grb2-DENND1A-Rab35 signaling pathway with the interaction of DENND1A and Grb2 as a regulatory center could regulate gastric cancer cell migration and invasion. Ultimately, the expression level of DENND1A predicts the survival status of gastric cancer patients and may become one of the important targets for the treatment of gastric cancer. |
| first_indexed | 2024-12-23T19:44:13Z |
| format | Article |
| id | doaj.art-700cc349d0c44c8e87ec8891ddad5156 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-23T19:44:13Z |
| publishDate | 2018-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-700cc349d0c44c8e87ec8891ddad51562022-12-21T17:33:35ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122018-11-01910.3389/fphar.2018.01343407091EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex FormationBixing Ye0Biao Duan1Wenjie Deng2Yueyuan Wang3Yan Chen4Jie Cui5Jie Cui6Shixiu Sun7Yujie Zhang8Yujie Zhang9Jun Du10Jun Du11Luo Gu12Luo Gu13Luo Gu14Lin Lin15Yurong Tang16Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaDepartment of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, ChinaDepartment of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Xuzhou Medical University, Xuzhou, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaJiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaJiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing Medical University, Nanjing, ChinaDepartment of Physiology, Nanjing Medical University, Nanjing, ChinaDepartment of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, ChinaJiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing Medical University, Nanjing, ChinaDepartment of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, ChinaDepartment of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, ChinaAims: The aim of this study was to reveal the specific molecular mechanisms by which DENND1A accepts EGF signaling and activates Rab35 in gastric cancer.Methods: The expression of proteins related to DENND1A was examined by western blot analysis. Activation of Rab35 was assessed by GST-pulldown. The interaction of DENND1A and Grb2 was assessed by GST-pulldown and co-immunoprecipitation assays. The relationship between DENND1A and cell migration and invasion was detected using wound healing and transwell by gene overexpression and RNA interference.Results: EGF stimulation significantly promoted cell migration, whereas transfection with siRab35 partially inhibited EGF-promoted cell migration. DENND1A is also involved in these processes and active Rab35. Moreover, DENND1A binds to the N-terminal and C-terminal SH3 domains of Grb2 through PRD. Of special interest is the observation that EGFR can recruit Grb2-DENND1A complex under EGF stimulation. Further results reveal that the higher the expression of DENND1A, the shorter progression-free survival of gastric cancer patients.Conclusion: In summary, we confirmed that EGF-Grb2-DENND1A-Rab35 signaling pathway with the interaction of DENND1A and Grb2 as a regulatory center could regulate gastric cancer cell migration and invasion. Ultimately, the expression level of DENND1A predicts the survival status of gastric cancer patients and may become one of the important targets for the treatment of gastric cancer.https://www.frontiersin.org/article/10.3389/fphar.2018.01343/fullRab35DENND1AGrb2interactioncell migrationgastric cancer |
| spellingShingle | Bixing Ye Biao Duan Wenjie Deng Yueyuan Wang Yan Chen Jie Cui Jie Cui Shixiu Sun Yujie Zhang Yujie Zhang Jun Du Jun Du Luo Gu Luo Gu Luo Gu Lin Lin Yurong Tang EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation Frontiers in Pharmacology Rab35 DENND1A Grb2 interaction cell migration gastric cancer |
| title | EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation |
| title_full | EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation |
| title_fullStr | EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation |
| title_full_unstemmed | EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation |
| title_short | EGF Stimulates Rab35 Activation and Gastric Cancer Cell Migration by Regulating DENND1A-Grb2 Complex Formation |
| title_sort | egf stimulates rab35 activation and gastric cancer cell migration by regulating dennd1a grb2 complex formation |
| topic | Rab35 DENND1A Grb2 interaction cell migration gastric cancer |
| url | https://www.frontiersin.org/article/10.3389/fphar.2018.01343/full |
| work_keys_str_mv | AT bixingye egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT biaoduan egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT wenjiedeng egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT yueyuanwang egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT yanchen egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT jiecui egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT jiecui egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT shixiusun egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT yujiezhang egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT yujiezhang egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT jundu egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT jundu egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT luogu egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT luogu egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT luogu egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT linlin egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation AT yurongtang egfstimulatesrab35activationandgastriccancercellmigrationbyregulatingdennd1agrb2complexformation |