RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma
Abstract Background Severe hypoxia is a prominent character of pancreatic ductal adenocarcinoma (PDAC) microenvironment. In the process of gemcitabine based chemotherapy, PDAC cells are insulted from replication stresses co-induced by hypoxia and gemcitabine. However, PDAC cells get outstanding abil...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-09-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-022-02490-3 |
| _version_ | 1798000651654922240 |
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| author | Qiu Tu Xiuyun Liu Xiaoqing Yao Ruixue Li Gaojing Liu Honglv Jiang Kaiqin Li Qiongfang Chen Xiaoyan Huang Qing Chang Guoqiang Xu Hong Zhu Peng Shi Bo Zhao |
| author_facet | Qiu Tu Xiuyun Liu Xiaoqing Yao Ruixue Li Gaojing Liu Honglv Jiang Kaiqin Li Qiongfang Chen Xiaoyan Huang Qing Chang Guoqiang Xu Hong Zhu Peng Shi Bo Zhao |
| author_sort | Qiu Tu |
| collection | DOAJ |
| description | Abstract Background Severe hypoxia is a prominent character of pancreatic ductal adenocarcinoma (PDAC) microenvironment. In the process of gemcitabine based chemotherapy, PDAC cells are insulted from replication stresses co-induced by hypoxia and gemcitabine. However, PDAC cells get outstanding abilities to resist to such harsh conditions and keep proliferating, causing a major obstacle for current therapy. RETSAT (Retinol Saturase) is defined as a hypoxia convergent gene recently, with high expression in PDAC hypoxic sectors. This study aimed to explore the roles of RETSAT in replication stress resistance and hypoxia adaptation in PDAC cells, and decipher the underlying mechanism. Methods The expression of RETSAT was examined in TCGA (The Cancer Genome Atlas), human pancreatic cancer microarray, clinical specimens and cell lines. Functions of RETSAT were studied by means of DNA fiber assay and comet assay in monolayer cultured PDAC cell lines, three dimensional spheroids, patient derived organoids and cell derived xenograft mouse models. Mechanism was investigated by using iPOND (isolate proteins on nascent DNA) combined with mass spectrometry, immunoprecipitation and immunoblotting. Results First, we found the converse relationship of RETSAT expression and PDAC chemotherapy. That is, PDAC patients with high RETSAT expression correlated with poor survival, while ones holding low RETSAT expression were benefitted more in Gemcitabine based chemotherapy. Second, we identified RETSAT as a novel replication fork associated protein. HIF-1α signaling promotes RETSAT expression under hypoxia. Functionally, RETSAT promoted fork restarting under replication stress and maintained genomic stability. Third, we uncovered the interaction of RETSAT and R-loop unwinding helicase DDX39B. RETSAT detained DDX39B on forks to resolve R-loops, through which avoided fork damage and CHK1 initiated apoptosis. Targeting DDX39B using chemical CCT018159 sensitized PDAC cells and organoids to gemcitabine induced apoptosis, highlighting the synergetic application of CCT018159 and gemcitabine in PDAC chemotherapy. Conclusions This study identified RETSAT as a novel replication fork protein, which functions through interacting with DDX39B mediated R-loop clearance to promote fork restarting, leading to cellular resistance to replication stresses co-induced by tumor environmental hypoxia and gemcitabine in pancreatic ductal adenocarcinoma. |
| first_indexed | 2024-04-11T11:23:44Z |
| format | Article |
| id | doaj.art-66e2c0f5f2b841ab9da4037fa74905fe |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-04-11T11:23:44Z |
| publishDate | 2022-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-66e2c0f5f2b841ab9da4037fa74905fe2022-12-22T04:26:21ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662022-09-0141112210.1186/s13046-022-02490-3RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinomaQiu Tu0Xiuyun Liu1Xiaoqing Yao2Ruixue Li3Gaojing Liu4Honglv Jiang5Kaiqin Li6Qiongfang Chen7Xiaoyan Huang8Qing Chang9Guoqiang Xu10Hong Zhu11Peng Shi12Bo Zhao13Kunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesDepartment of Hepatobiliary and Pancreatic Surgery, Second Affiliated Hospital of Kunming Medical UniversityKunming Institute of Zoology, Chinese Academy of SciencesJiangsu Key Laboratory of Neuropsychiatric Diseases and College of Pharmaceutical Sciences, Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow UniversityKunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesJiangsu Key Laboratory of Neuropsychiatric Diseases and College of Pharmaceutical Sciences, Jiangsu Key Laboratory of Preventive and Translational Medicine for Geriatric Diseases, Soochow UniversityDepartment of Hepatobiliary and Pancreatic Surgery, Second Affiliated Hospital of Kunming Medical UniversityKunming Institute of Zoology, Chinese Academy of SciencesKunming Institute of Zoology, Chinese Academy of SciencesAbstract Background Severe hypoxia is a prominent character of pancreatic ductal adenocarcinoma (PDAC) microenvironment. In the process of gemcitabine based chemotherapy, PDAC cells are insulted from replication stresses co-induced by hypoxia and gemcitabine. However, PDAC cells get outstanding abilities to resist to such harsh conditions and keep proliferating, causing a major obstacle for current therapy. RETSAT (Retinol Saturase) is defined as a hypoxia convergent gene recently, with high expression in PDAC hypoxic sectors. This study aimed to explore the roles of RETSAT in replication stress resistance and hypoxia adaptation in PDAC cells, and decipher the underlying mechanism. Methods The expression of RETSAT was examined in TCGA (The Cancer Genome Atlas), human pancreatic cancer microarray, clinical specimens and cell lines. Functions of RETSAT were studied by means of DNA fiber assay and comet assay in monolayer cultured PDAC cell lines, three dimensional spheroids, patient derived organoids and cell derived xenograft mouse models. Mechanism was investigated by using iPOND (isolate proteins on nascent DNA) combined with mass spectrometry, immunoprecipitation and immunoblotting. Results First, we found the converse relationship of RETSAT expression and PDAC chemotherapy. That is, PDAC patients with high RETSAT expression correlated with poor survival, while ones holding low RETSAT expression were benefitted more in Gemcitabine based chemotherapy. Second, we identified RETSAT as a novel replication fork associated protein. HIF-1α signaling promotes RETSAT expression under hypoxia. Functionally, RETSAT promoted fork restarting under replication stress and maintained genomic stability. Third, we uncovered the interaction of RETSAT and R-loop unwinding helicase DDX39B. RETSAT detained DDX39B on forks to resolve R-loops, through which avoided fork damage and CHK1 initiated apoptosis. Targeting DDX39B using chemical CCT018159 sensitized PDAC cells and organoids to gemcitabine induced apoptosis, highlighting the synergetic application of CCT018159 and gemcitabine in PDAC chemotherapy. Conclusions This study identified RETSAT as a novel replication fork protein, which functions through interacting with DDX39B mediated R-loop clearance to promote fork restarting, leading to cellular resistance to replication stresses co-induced by tumor environmental hypoxia and gemcitabine in pancreatic ductal adenocarcinoma.https://doi.org/10.1186/s13046-022-02490-3RETSATDDX39BFork restartingHypoxiaGemcitabineResistance |
| spellingShingle | Qiu Tu Xiuyun Liu Xiaoqing Yao Ruixue Li Gaojing Liu Honglv Jiang Kaiqin Li Qiongfang Chen Xiaoyan Huang Qing Chang Guoqiang Xu Hong Zhu Peng Shi Bo Zhao RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma Journal of Experimental & Clinical Cancer Research RETSAT DDX39B Fork restarting Hypoxia Gemcitabine Resistance |
| title | RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| title_full | RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| title_fullStr | RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| title_full_unstemmed | RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| title_short | RETSAT associates with DDX39B to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| title_sort | retsat associates with ddx39b to promote fork restarting and resistance to gemcitabine based chemotherapy in pancreatic ductal adenocarcinoma |
| topic | RETSAT DDX39B Fork restarting Hypoxia Gemcitabine Resistance |
| url | https://doi.org/10.1186/s13046-022-02490-3 |
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