Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models
Abstract Cancer stem cells (CSCs) are present in small quantities in tumor populations. To permit various analyses of CSCs, we attempted to enrich and expand ornithine decarboxylase (ODC) degron-transduced colorectal cancer (CRC) cells, which retain low proteasome activity. ZsGreen fluorescence-posi...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-025-04277-w |
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| author | Ryo Ikeshima Hidekazu Takahashi Hiroyuki Yamamoto Shihori Kouda Rika Akisue Manami Tsujimoto Yuhki Yokoyama Haruka Hirose Hiroaki Itakura Yoshihiro Morimoto Norikatsu Miyoshi Mamoru Uemura Daisuke Okuzaki Shinji Tanaka Hidetoshi Eguchi Yuichiro Doki Hirofumi Yamamoto Masaki Mori |
| author_facet | Ryo Ikeshima Hidekazu Takahashi Hiroyuki Yamamoto Shihori Kouda Rika Akisue Manami Tsujimoto Yuhki Yokoyama Haruka Hirose Hiroaki Itakura Yoshihiro Morimoto Norikatsu Miyoshi Mamoru Uemura Daisuke Okuzaki Shinji Tanaka Hidetoshi Eguchi Yuichiro Doki Hirofumi Yamamoto Masaki Mori |
| author_sort | Ryo Ikeshima |
| collection | DOAJ |
| description | Abstract Cancer stem cells (CSCs) are present in small quantities in tumor populations. To permit various analyses of CSCs, we attempted to enrich and expand ornithine decarboxylase (ODC) degron-transduced colorectal cancer (CRC) cells, which retain low proteasome activity. ZsGreen fluorescence-positive (ZsGreen+) cells were collected by sorting the ODC degron-transduced HCT116, DLD1, and SW480 cells, which were defined as enriched ZsGreen+ cells. ZsGreen+ cells still maintained CSC properties. These cells had higher stem cell marker expression and increased resistance to chemotherapy with 5-fluorouracil and oxaliplatin. ZsGreen+ HCT116 and DLD1 cells had greater sphere-forming ability and enhanced tumorigenicity compared to ZsGreen- control cells. Time-lapse microscopy showed that a single enriched ZsGreen+ HCT116 cell had asymmetric cell division. Thus, cancer stem model cells were acquired in sufficient quantity. Using these cells, we performed a comprehensive microRNA analysis; miR-491-3p was a candidate to suppress cancer stemness. Finally, we found that up-regulated genes in the enriched HCT116 ZsGreen+ cells correlated with those up-regulated in human clinical spheroid samples established from patient-derived xenografts (PDXs) derived from CRC tissue samples, further supporting the acquisition of enriched model CSCs. These cells with the aid of clinical spheroid samples would be useful in identifying novel CSC markers and developing medicine for anti-CSC therapy. |
| format | Article |
| id | doaj-art-e3d4263a6a9a4653a0fee5f791dd803e |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-e3d4263a6a9a4653a0fee5f791dd803e2025-08-20T03:37:29ZengNature PortfolioScientific Reports2045-23222025-07-0115111610.1038/s41598-025-04277-wEnrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell modelsRyo Ikeshima0Hidekazu Takahashi1Hiroyuki Yamamoto2Shihori Kouda3Rika Akisue4Manami Tsujimoto5Yuhki Yokoyama6Haruka Hirose7Hiroaki Itakura8Yoshihiro Morimoto9Norikatsu Miyoshi10Mamoru Uemura11Daisuke Okuzaki12Shinji Tanaka13Hidetoshi Eguchi14Yuichiro Doki15Hirofumi Yamamoto16Masaki Mori17Department of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Molecular Pathology, Division of Health Sciences, Graduate School of Medicine, Osaka UniversityDepartment of Molecular Pathology, Division of Health Sciences, Graduate School of Medicine, Osaka UniversityDepartment of Molecular Pathology, Division of Health Sciences, Graduate School of Medicine, Osaka UniversityDepartment of Molecular Pathology, Division of Health Sciences, Graduate School of Medicine, Osaka UniversityDepartment of Molecular Pathology, Division of Health Sciences, Graduate School of Medicine, Osaka UniversityDepartment of Computational and Systems Biology, Medical Research Institute, Institute of Science TokyoDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityGenome Information Research Centre, Research Institute for Microbial Diseases, Osaka UniversityDepartment of Molecular Oncology, Graduate School of Medicine, Institute of Science TokyoDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityDepartment of Surgery, Gastroenterological Surgery, Graduate School of Medicine, Osaka UniversityGraduate School of Medicine, Tokai UniversityAbstract Cancer stem cells (CSCs) are present in small quantities in tumor populations. To permit various analyses of CSCs, we attempted to enrich and expand ornithine decarboxylase (ODC) degron-transduced colorectal cancer (CRC) cells, which retain low proteasome activity. ZsGreen fluorescence-positive (ZsGreen+) cells were collected by sorting the ODC degron-transduced HCT116, DLD1, and SW480 cells, which were defined as enriched ZsGreen+ cells. ZsGreen+ cells still maintained CSC properties. These cells had higher stem cell marker expression and increased resistance to chemotherapy with 5-fluorouracil and oxaliplatin. ZsGreen+ HCT116 and DLD1 cells had greater sphere-forming ability and enhanced tumorigenicity compared to ZsGreen- control cells. Time-lapse microscopy showed that a single enriched ZsGreen+ HCT116 cell had asymmetric cell division. Thus, cancer stem model cells were acquired in sufficient quantity. Using these cells, we performed a comprehensive microRNA analysis; miR-491-3p was a candidate to suppress cancer stemness. Finally, we found that up-regulated genes in the enriched HCT116 ZsGreen+ cells correlated with those up-regulated in human clinical spheroid samples established from patient-derived xenografts (PDXs) derived from CRC tissue samples, further supporting the acquisition of enriched model CSCs. These cells with the aid of clinical spheroid samples would be useful in identifying novel CSC markers and developing medicine for anti-CSC therapy.https://doi.org/10.1038/s41598-025-04277-w |
| spellingShingle | Ryo Ikeshima Hidekazu Takahashi Hiroyuki Yamamoto Shihori Kouda Rika Akisue Manami Tsujimoto Yuhki Yokoyama Haruka Hirose Hiroaki Itakura Yoshihiro Morimoto Norikatsu Miyoshi Mamoru Uemura Daisuke Okuzaki Shinji Tanaka Hidetoshi Eguchi Yuichiro Doki Hirofumi Yamamoto Masaki Mori Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models Scientific Reports |
| title | Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| title_full | Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| title_fullStr | Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| title_full_unstemmed | Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| title_short | Enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| title_sort | enrichment of ornithine decarboxylase degron transduced colorectal cancer cells for extended application of cancer stem cell models |
| url | https://doi.org/10.1038/s41598-025-04277-w |
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