Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer

Abstract Purpose Radiation therapy has revolutionized the treatment of primary or liver metastases in colorectal cancer (CRC). In colorectal cancer, conventional fractionation (1.8 ~ 2.0 Gy daily) is typically used for treatment. Nevertheless, there is a paucity of research investigating the potenti...

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Main Authors: Shuxuan Wang, Yanyan Lin, Qianqian Zhao, Huanliang Chen, Shisuo Du, Zhaochong Zeng
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Molecular Medicine
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Online Access:https://doi.org/10.1186/s10020-025-01206-5
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author Shuxuan Wang
Yanyan Lin
Qianqian Zhao
Huanliang Chen
Shisuo Du
Zhaochong Zeng
author_facet Shuxuan Wang
Yanyan Lin
Qianqian Zhao
Huanliang Chen
Shisuo Du
Zhaochong Zeng
author_sort Shuxuan Wang
collection DOAJ
description Abstract Purpose Radiation therapy has revolutionized the treatment of primary or liver metastases in colorectal cancer (CRC). In colorectal cancer, conventional fractionation (1.8 ~ 2.0 Gy daily) is typically used for treatment. Nevertheless, there is a paucity of research investigating the potential implications of radiation therapy-induced alterations in the expression levels of regulatory genes on resistance to chemotherapy agents. Herein, we explored the mechanism by which conventional fractionation drives 5-fluorouracil (5-FU) resistance and metformin (Met) rescued 5-FU resistance in CRC. Methods and materials RNA sequencing, differential genes expression analysis was performed to identify the 5-FU resistance genes after irradiation (according to the convention of cell irradiation, 2 Gy × 8 scheme was selected). Drug sensitivity assay, immunofluorescence staining, folate analogs concentration measurement was used to explore the biological function of histocompatibility minor 13 (HM13) and γ-Glutamyl Hydrolase (GGH). Combined chemosensitivity test and xenograft mouse model has been used to gain insights into the underlying clinical value of the combination of 5-FU and Met. Results The conventional fractionation scheme (2 Gy × 8) induced resistance to 5-FU in the CRC cell line HCT-15, accompanied by an elevated RNA expression level of peptidase HM13. Mechanistically, the increased expression of HM13 caused an abnormal shearing of the N-terminal signal peptide of γ-Glutamyl Hydrolase (GGH), which resulted in decreased intracellular content of 5, 10-methylenetetrahydrofolate (5,10-CH2-THF). Conclusion We revealed a new mechanism of 5-FU resistance induced by irradiated with 2 Gy × 8 through the HM13-GGH-5,10-CH2-THF axis. The synergistic effect of Met and 5-FU can rescue 5-FU resistance after conventional fractionated irradiation. In summary, this work will help to reveal the mechanisms of IR-induced 5-FU resistance, which is important for finding new therapeutic targets and improving the efficacy of chemotherapy regimens after radiotherapy.
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spelling doaj-art-c7b1599f95ea46b5acb7929f2c5f3a392025-08-20T02:33:25ZengBMCMolecular Medicine1528-36582025-05-0131111510.1186/s10020-025-01206-5Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancerShuxuan Wang0Yanyan Lin1Qianqian Zhao2Huanliang Chen3Shisuo Du4Zhaochong Zeng5Department of Radiation Oncology, Zhongshan Hospital, Fudan UniversityDepartment of Radiation Oncology, Zhongshan Hospital, Fudan UniversityDepartment of Radiation Oncology, Zhongshan Hospital, Fudan UniversityDepartment of Radiation Oncology, Zhongshan Hospital, Fudan UniversityDepartment of Radiation Oncology, Zhongshan Hospital, Fudan UniversityDepartment of Radiation Oncology, Zhongshan Hospital, Fudan UniversityAbstract Purpose Radiation therapy has revolutionized the treatment of primary or liver metastases in colorectal cancer (CRC). In colorectal cancer, conventional fractionation (1.8 ~ 2.0 Gy daily) is typically used for treatment. Nevertheless, there is a paucity of research investigating the potential implications of radiation therapy-induced alterations in the expression levels of regulatory genes on resistance to chemotherapy agents. Herein, we explored the mechanism by which conventional fractionation drives 5-fluorouracil (5-FU) resistance and metformin (Met) rescued 5-FU resistance in CRC. Methods and materials RNA sequencing, differential genes expression analysis was performed to identify the 5-FU resistance genes after irradiation (according to the convention of cell irradiation, 2 Gy × 8 scheme was selected). Drug sensitivity assay, immunofluorescence staining, folate analogs concentration measurement was used to explore the biological function of histocompatibility minor 13 (HM13) and γ-Glutamyl Hydrolase (GGH). Combined chemosensitivity test and xenograft mouse model has been used to gain insights into the underlying clinical value of the combination of 5-FU and Met. Results The conventional fractionation scheme (2 Gy × 8) induced resistance to 5-FU in the CRC cell line HCT-15, accompanied by an elevated RNA expression level of peptidase HM13. Mechanistically, the increased expression of HM13 caused an abnormal shearing of the N-terminal signal peptide of γ-Glutamyl Hydrolase (GGH), which resulted in decreased intracellular content of 5, 10-methylenetetrahydrofolate (5,10-CH2-THF). Conclusion We revealed a new mechanism of 5-FU resistance induced by irradiated with 2 Gy × 8 through the HM13-GGH-5,10-CH2-THF axis. The synergistic effect of Met and 5-FU can rescue 5-FU resistance after conventional fractionated irradiation. In summary, this work will help to reveal the mechanisms of IR-induced 5-FU resistance, which is important for finding new therapeutic targets and improving the efficacy of chemotherapy regimens after radiotherapy.https://doi.org/10.1186/s10020-025-01206-5Irradiation5-FUHistocompatibility minor 13MetforminFolate metabolism
spellingShingle Shuxuan Wang
Yanyan Lin
Qianqian Zhao
Huanliang Chen
Shisuo Du
Zhaochong Zeng
Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
Molecular Medicine
Irradiation
5-FU
Histocompatibility minor 13
Metformin
Folate metabolism
title Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
title_full Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
title_fullStr Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
title_full_unstemmed Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
title_short Metformin reverses 5-FU resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
title_sort metformin reverses 5 fu resistance induced by radiotherapy through mediating folate metabolism in colorectal cancer
topic Irradiation
5-FU
Histocompatibility minor 13
Metformin
Folate metabolism
url https://doi.org/10.1186/s10020-025-01206-5
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