MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer
Abstract Oxaliplatin (Oxa) has been extensively employed in treatment of colorectal cancer (CRC), yet frequent occurrence of chemoresistance poses a significant obstacle to achieving long-term disease-free survival for CRC patients. In this study, we employed a CRISPR/Cas9 sgRNA library targeting 1,...
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2025-07-01
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| Online Access: | https://doi.org/10.1186/s10020-025-01287-2 |
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| author | Yixuan Wang Mu Qiao Jing Guo Ying Xie Meilin Hu Xin Li Sheng Wang Jingjing Wang Jingya Wang Ziyi Peng Mengqi Wang Hao Cheng Tiantian Li Linchuang Jia Danchen Su Huanhuan Liu Kexin Hu Xinyang Li Wenjing Li Di Wu Zhe Zhang Jianing Han Ruiyang Bai Funan Zhou Zhiqiang Liu |
| author_facet | Yixuan Wang Mu Qiao Jing Guo Ying Xie Meilin Hu Xin Li Sheng Wang Jingjing Wang Jingya Wang Ziyi Peng Mengqi Wang Hao Cheng Tiantian Li Linchuang Jia Danchen Su Huanhuan Liu Kexin Hu Xinyang Li Wenjing Li Di Wu Zhe Zhang Jianing Han Ruiyang Bai Funan Zhou Zhiqiang Liu |
| author_sort | Yixuan Wang |
| collection | DOAJ |
| description | Abstract Oxaliplatin (Oxa) has been extensively employed in treatment of colorectal cancer (CRC), yet frequent occurrence of chemoresistance poses a significant obstacle to achieving long-term disease-free survival for CRC patients. In this study, we employed a CRISPR/Cas9 sgRNA library targeting 1,117 human ubiquitination-related genes to screen key regulators of sensitivity to Oxa and identified the Makorin Ring Finger Protein 1 (MKRN1), an E3 ligase, as an Oxa-resistant gene of CRC cells. Clinically, MKRN1 is highly expressed in CRC tissues compared with the adjacent normal tissue, and its upregulation is correlated with poor therapeutic response, disease progression, and worse overall survival of CRC patients treated with Oxa-based regimens. In CRC cells, gain- and loss-of-function studies of MKRN1 respectively altered the sensitivity to Oxa treatment, as evidenced by changes in IC50 values and cell apoptosis. Mechanistic analysis revealed that MKRN1 interacts with Aspartate/Glutamate Carrier 1 (AGC1), facilitating degradation of AGC1 via K11- and K29-linked ubiquitination, thereby affecting mitochondrial function including energy metabolism and antioxidant responses. Through reprogramming of metabolic genes, this process enhances the expression of heat shock protein HSPD1 and HSP90AA1, while reducing oxidative stress, ultimately contributing to the development of Oxa resistance in CRC cells. Moreover, AGC1 knockdown rescued the MKRN1-deficiency induced Oxa-sensitivity of CRC cells and in xenograft mouse model. Translationally, we identified Rabdosiin as a potential inhibitor of MKRN1 through virtual screening, and validated the synergetic effect of Rabdosiin and Oxa in treating Oxa-resistant CRC cells both in vitro and in vivo. Taken together, our findings highlight the pivotal role of MKRN1-AGC1 axis in dictating CRC chemoresistance and offer novel therapeutic strategies for overcoming Oxa-resistance. Graphical Abstract |
| format | Article |
| id | doaj-art-49937ca09a674536b8641cf1be92a26f |
| institution | DOAJ |
| issn | 1528-3658 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
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| series | Molecular Medicine |
| spelling | doaj-art-49937ca09a674536b8641cf1be92a26f2025-08-20T03:05:04ZengBMCMolecular Medicine1528-36582025-07-0131111410.1186/s10020-025-01287-2MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal CancerYixuan Wang0Mu Qiao1Jing Guo2Ying Xie3Meilin Hu4Xin Li5Sheng Wang6Jingjing Wang7Jingya Wang8Ziyi Peng9Mengqi Wang10Hao Cheng11Tiantian Li12Linchuang Jia13Danchen Su14Huanhuan Liu15Kexin Hu16Xinyang Li17Wenjing Li18Di Wu19Zhe Zhang20Jianing Han21Ruiyang Bai22Funan Zhou23Zhiqiang Liu24The province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesTianjin Key Laboratory of Oral Soft and Hard Tissues Restoration and Regeneration, School of stomatology, Tianjin Medical University, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesDepartment of Hematology, Tianjin Medical University Cancer Hospital, Tianjin Medical UniversityShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesShandong Provincial Key Laboratory of Precision Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University, Shandong Academy of Medical SciencesThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityThe province and ministry co-sponsored collaborative innovation center for medical epigenetics, Department of Physiology and Pathophysiology, School of Basic Medical Science, Tianjin Medical UniversityAbstract Oxaliplatin (Oxa) has been extensively employed in treatment of colorectal cancer (CRC), yet frequent occurrence of chemoresistance poses a significant obstacle to achieving long-term disease-free survival for CRC patients. In this study, we employed a CRISPR/Cas9 sgRNA library targeting 1,117 human ubiquitination-related genes to screen key regulators of sensitivity to Oxa and identified the Makorin Ring Finger Protein 1 (MKRN1), an E3 ligase, as an Oxa-resistant gene of CRC cells. Clinically, MKRN1 is highly expressed in CRC tissues compared with the adjacent normal tissue, and its upregulation is correlated with poor therapeutic response, disease progression, and worse overall survival of CRC patients treated with Oxa-based regimens. In CRC cells, gain- and loss-of-function studies of MKRN1 respectively altered the sensitivity to Oxa treatment, as evidenced by changes in IC50 values and cell apoptosis. Mechanistic analysis revealed that MKRN1 interacts with Aspartate/Glutamate Carrier 1 (AGC1), facilitating degradation of AGC1 via K11- and K29-linked ubiquitination, thereby affecting mitochondrial function including energy metabolism and antioxidant responses. Through reprogramming of metabolic genes, this process enhances the expression of heat shock protein HSPD1 and HSP90AA1, while reducing oxidative stress, ultimately contributing to the development of Oxa resistance in CRC cells. Moreover, AGC1 knockdown rescued the MKRN1-deficiency induced Oxa-sensitivity of CRC cells and in xenograft mouse model. Translationally, we identified Rabdosiin as a potential inhibitor of MKRN1 through virtual screening, and validated the synergetic effect of Rabdosiin and Oxa in treating Oxa-resistant CRC cells both in vitro and in vivo. Taken together, our findings highlight the pivotal role of MKRN1-AGC1 axis in dictating CRC chemoresistance and offer novel therapeutic strategies for overcoming Oxa-resistance. Graphical Abstracthttps://doi.org/10.1186/s10020-025-01287-2Colorectal cancerUbiquitinationMKRN1AGC1Heat shock proteinsMitochondrial energy metabolism |
| spellingShingle | Yixuan Wang Mu Qiao Jing Guo Ying Xie Meilin Hu Xin Li Sheng Wang Jingjing Wang Jingya Wang Ziyi Peng Mengqi Wang Hao Cheng Tiantian Li Linchuang Jia Danchen Su Huanhuan Liu Kexin Hu Xinyang Li Wenjing Li Di Wu Zhe Zhang Jianing Han Ruiyang Bai Funan Zhou Zhiqiang Liu MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer Molecular Medicine Colorectal cancer Ubiquitination MKRN1 AGC1 Heat shock proteins Mitochondrial energy metabolism |
| title | MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer |
| title_full | MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer |
| title_fullStr | MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer |
| title_full_unstemmed | MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer |
| title_short | MKRN1 degrades AGC1 to trigger chemotherapy resistance of colorectal Cancer |
| title_sort | mkrn1 degrades agc1 to trigger chemotherapy resistance of colorectal cancer |
| topic | Colorectal cancer Ubiquitination MKRN1 AGC1 Heat shock proteins Mitochondrial energy metabolism |
| url | https://doi.org/10.1186/s10020-025-01287-2 |
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