m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway
Abstract Background Gemcitabine resistance has brought great challenges to the treatment of pancreatic cancer. The N6-methyladenosine (m6A) mutation has been shown to have a significant regulatory role in chemosensitivity; however, it is not apparent whether gemcitabine resistance can be regulated b...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-08-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-023-02792-0 |
| _version_ | 1797451076211834880 |
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| author | Kai Lin Endi Zhou Ting Shi Siqing Zhang Jinfan Zhang Ziruo Zheng Yuetian Pan Wentao Gao Yabin Yu |
| author_facet | Kai Lin Endi Zhou Ting Shi Siqing Zhang Jinfan Zhang Ziruo Zheng Yuetian Pan Wentao Gao Yabin Yu |
| author_sort | Kai Lin |
| collection | DOAJ |
| description | Abstract Background Gemcitabine resistance has brought great challenges to the treatment of pancreatic cancer. The N6-methyladenosine (m6A) mutation has been shown to have a significant regulatory role in chemosensitivity; however, it is not apparent whether gemcitabine resistance can be regulated by fat mass and obesity-associated protein (FTO). Methods Cells with established gemcitabine resistance and tissues from pancreatic cancer patients were used to evaluate FTO expression. The biological mechanisms of the effects of FTO on gemcitabine resistant cells were investigated using CCK-8, colony formation assay, flow cytometry, and inhibitory concentration 50. Immunoprecipitation/mass spectrometry, MeRIP-seq, RNA sequencing and RIP assays, RNA stability, luciferase reporter, and RNA pull down assays were employed to examine the mechanism of FTO affecting gemcitabine resistant pancreatic cancer cells. Results The results revealed that FTO was substantially expressed in cells and tissues that were resistant to gemcitabine. Functionally, the gemcitabine resistance of pancreatic cancer could be enhanced by FTO, while its depletion inhibited the growth of gemcitabine resistant tumor cells in vivo. Immunoprecipitation/mass spectrometry showed that the FTO protein can be bound to USP7 and deubiquitinated by USP7, leading to the upregulation of FTO. At the same time, FTO knockdown significantly decreased the expression level of NEDD4 in an m6A-dependent manner. RNA pull down and RNA immunoprecipitation verified YTHDF2 as the reader of NEDD4, which promoted the chemoresistance of gemcitabine resistant cells. FTO knockdown markedly increased the PTEN expression level in an NEDD4-dependent manner and influenced the chemosensitivity to gemcitabine through the PI3K/AKT pathway in pancreatic cancer cells. Conclusion In conclusion, we found that gemcitabine resistance in pancreatic cancer can be influenced by FTO that demethylates NEDD4 RNA in a m6A-dependent manner, which then influences the PTEN expression level and thereby affects the PI3K/AKT pathway. We also identified that the FTO level can be upregulated by USP7. |
| first_indexed | 2024-03-09T14:49:44Z |
| format | Article |
| id | doaj.art-d556c8e2e35c4b8aa19b1183d4681c03 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-03-09T14:49:44Z |
| publishDate | 2023-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-d556c8e2e35c4b8aa19b1183d4681c032023-11-26T14:34:37ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662023-08-0142111910.1186/s13046-023-02792-0m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathwayKai Lin0Endi Zhou1Ting Shi2Siqing Zhang3Jinfan Zhang4Ziruo Zheng5Yuetian Pan6Wentao Gao7Yabin Yu8Department of Gastrointestinal Surgery, Affiliated Hangzhou First People’s Hospital, Zhejiang University School of MedicinePancreas Center, the First Affiliated Hospital of Nanjing Medical UniversityDepartment of Hepatobiliary Surgery, The Afliated Huaian No.1 People’s Hospital of Nanjing Medical UniversityPancreas Center, the First Affiliated Hospital of Nanjing Medical UniversityPancreas Center, the First Affiliated Hospital of Nanjing Medical UniversityPancreas Center, the First Affiliated Hospital of Nanjing Medical UniversityMedical Faculty of Ludwig Maximilians, University of Munich-MunichPancreas Center, the First Affiliated Hospital of Nanjing Medical UniversityDepartment of Hepatobiliary Surgery, The Afliated Huaian No.1 People’s Hospital of Nanjing Medical UniversityAbstract Background Gemcitabine resistance has brought great challenges to the treatment of pancreatic cancer. The N6-methyladenosine (m6A) mutation has been shown to have a significant regulatory role in chemosensitivity; however, it is not apparent whether gemcitabine resistance can be regulated by fat mass and obesity-associated protein (FTO). Methods Cells with established gemcitabine resistance and tissues from pancreatic cancer patients were used to evaluate FTO expression. The biological mechanisms of the effects of FTO on gemcitabine resistant cells were investigated using CCK-8, colony formation assay, flow cytometry, and inhibitory concentration 50. Immunoprecipitation/mass spectrometry, MeRIP-seq, RNA sequencing and RIP assays, RNA stability, luciferase reporter, and RNA pull down assays were employed to examine the mechanism of FTO affecting gemcitabine resistant pancreatic cancer cells. Results The results revealed that FTO was substantially expressed in cells and tissues that were resistant to gemcitabine. Functionally, the gemcitabine resistance of pancreatic cancer could be enhanced by FTO, while its depletion inhibited the growth of gemcitabine resistant tumor cells in vivo. Immunoprecipitation/mass spectrometry showed that the FTO protein can be bound to USP7 and deubiquitinated by USP7, leading to the upregulation of FTO. At the same time, FTO knockdown significantly decreased the expression level of NEDD4 in an m6A-dependent manner. RNA pull down and RNA immunoprecipitation verified YTHDF2 as the reader of NEDD4, which promoted the chemoresistance of gemcitabine resistant cells. FTO knockdown markedly increased the PTEN expression level in an NEDD4-dependent manner and influenced the chemosensitivity to gemcitabine through the PI3K/AKT pathway in pancreatic cancer cells. Conclusion In conclusion, we found that gemcitabine resistance in pancreatic cancer can be influenced by FTO that demethylates NEDD4 RNA in a m6A-dependent manner, which then influences the PTEN expression level and thereby affects the PI3K/AKT pathway. We also identified that the FTO level can be upregulated by USP7.https://doi.org/10.1186/s13046-023-02792-0FTON6-methyladenosinePancreatic carcinomaProliferationGemcitabine resistance |
| spellingShingle | Kai Lin Endi Zhou Ting Shi Siqing Zhang Jinfan Zhang Ziruo Zheng Yuetian Pan Wentao Gao Yabin Yu m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway Journal of Experimental & Clinical Cancer Research FTO N6-methyladenosine Pancreatic carcinoma Proliferation Gemcitabine resistance |
| title | m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway |
| title_full | m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway |
| title_fullStr | m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway |
| title_full_unstemmed | m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway |
| title_short | m6A eraser FTO impairs gemcitabine resistance in pancreatic cancer through influencing NEDD4 mRNA stability by regulating the PTEN/PI3K/AKT pathway |
| title_sort | m6a eraser fto impairs gemcitabine resistance in pancreatic cancer through influencing nedd4 mrna stability by regulating the pten pi3k akt pathway |
| topic | FTO N6-methyladenosine Pancreatic carcinoma Proliferation Gemcitabine resistance |
| url | https://doi.org/10.1186/s13046-023-02792-0 |
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