An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids
Abstract Background Ovarian cancer (OC) is the most lethal gynecological cancer due to the recurrence of drug-resistance. Cancer initiating cells (CICs) are proposed to be responsible for the aggressiveness of OC. The rarity and difficulty of in vitro long-term cultivation of CICs challenge the deve...
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| Format: | Article |
| Language: | English |
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BMC
2022-09-01
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| Series: | BMC Cancer |
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| Online Access: | https://doi.org/10.1186/s12885-022-10042-3 |
| _version_ | 1811273563848572928 |
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| author | Yu-An Chen Chen-Yu Lu Wen-Fang Cheng Kuan-Ting Kuo Chen-Wei Yu Hon-Nerng Ho Hsin-Fu Chen Szu-Hua Pan |
| author_facet | Yu-An Chen Chen-Yu Lu Wen-Fang Cheng Kuan-Ting Kuo Chen-Wei Yu Hon-Nerng Ho Hsin-Fu Chen Szu-Hua Pan |
| author_sort | Yu-An Chen |
| collection | DOAJ |
| description | Abstract Background Ovarian cancer (OC) is the most lethal gynecological cancer due to the recurrence of drug-resistance. Cancer initiating cells (CICs) are proposed to be responsible for the aggressiveness of OC. The rarity and difficulty of in vitro long-term cultivation of CICs challenge the development of CIC-targeting therapeutics. Reprogramming cancer cells into induced cancer initiating cell (iCICs) could be an approach to solve these. Several inducible CICs have been acquired by activating the expression of stemness genes in different cancer cells. However, few reports have demonstrated the feasibility in OC. Methods Patients with primary OC receiving surgery were enrolled. Tumor tissue were collected, and OCT4, SOX2, and NANOG expressions were assessed by immunohistochemistry (IHC) staining to investigate the association of stemness markers with overall survival (OS). An high-grade serous ovarian cancer (HGSOC) cell line, OVCAR-3 was reprogrammed by transducing Yamanaka four factors OCT4, SOX2, KLF4 and MYC (OSKM) to establish an iOCIC model, iOVCAR-3-OSKM. CIC characteristics of iOVCAR-3-OSKM were evaluated by RT-PCR, sphere formation assay and animal experiments. Drug-resistance and migration ability were accessed by dye-efflux activity assay, MTT assay and migration assay. Gene profile was presented through RNA-sequencing. Lineage differentiation ability and organoid culture were determined by in vitro differentiation assays. Results In OC patients, the co-expression of multiple stem-related transcription factors (OCT4, SOX2, and NANOG) was associated with worse OS. iOVCAR-3-OSKM cells generated by reprogramming successfully exhibited stemness characteristics with strong sphere-forming and tumorigenesis ability. iOVCAR-3-OSKM cells also showed malignant potential with higher drug resistance to chemodrug, Paclitaxel (PTX) and migration ability. iOVCAR-3-OSKM was maintainable and expandable on feeder-dependent culture condition, it also preserved ovarian lineage differentiation abilities, which could well differentiate into OC cells with CK-7 and CA125 expressions and develop into an organoid mimic poor prognostic OC histological feature. Conclusions The establishment of iOVCAR-3-OSKM not only allows us to fill the gap in the information on induced CICs in OC but also provides a potential strategy to develop personalized CICs and organoid models for treating OC in the near future. |
| first_indexed | 2024-04-12T23:01:43Z |
| format | Article |
| id | doaj.art-d640bfdf68c841e889c2d284c10295bc |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-04-12T23:01:43Z |
| publishDate | 2022-09-01 |
| publisher | BMC |
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| series | BMC Cancer |
| spelling | doaj.art-d640bfdf68c841e889c2d284c10295bc2022-12-22T03:13:03ZengBMCBMC Cancer1471-24072022-09-0122112110.1186/s12885-022-10042-3An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoidsYu-An Chen0Chen-Yu Lu1Wen-Fang Cheng2Kuan-Ting Kuo3Chen-Wei Yu4Hon-Nerng Ho5Hsin-Fu Chen6Szu-Hua Pan7Graduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityGraduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityGraduate Institute of Oncology, College of Medicine, National Taiwan UniversityDepartment of Pathology and Graduate Institute of Pathology, College of Medicine, National Taiwan UniversityGraduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityGraduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityGraduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityGraduate Institute of Medical Genomics and Proteomics, College of Medicine, National Taiwan UniversityAbstract Background Ovarian cancer (OC) is the most lethal gynecological cancer due to the recurrence of drug-resistance. Cancer initiating cells (CICs) are proposed to be responsible for the aggressiveness of OC. The rarity and difficulty of in vitro long-term cultivation of CICs challenge the development of CIC-targeting therapeutics. Reprogramming cancer cells into induced cancer initiating cell (iCICs) could be an approach to solve these. Several inducible CICs have been acquired by activating the expression of stemness genes in different cancer cells. However, few reports have demonstrated the feasibility in OC. Methods Patients with primary OC receiving surgery were enrolled. Tumor tissue were collected, and OCT4, SOX2, and NANOG expressions were assessed by immunohistochemistry (IHC) staining to investigate the association of stemness markers with overall survival (OS). An high-grade serous ovarian cancer (HGSOC) cell line, OVCAR-3 was reprogrammed by transducing Yamanaka four factors OCT4, SOX2, KLF4 and MYC (OSKM) to establish an iOCIC model, iOVCAR-3-OSKM. CIC characteristics of iOVCAR-3-OSKM were evaluated by RT-PCR, sphere formation assay and animal experiments. Drug-resistance and migration ability were accessed by dye-efflux activity assay, MTT assay and migration assay. Gene profile was presented through RNA-sequencing. Lineage differentiation ability and organoid culture were determined by in vitro differentiation assays. Results In OC patients, the co-expression of multiple stem-related transcription factors (OCT4, SOX2, and NANOG) was associated with worse OS. iOVCAR-3-OSKM cells generated by reprogramming successfully exhibited stemness characteristics with strong sphere-forming and tumorigenesis ability. iOVCAR-3-OSKM cells also showed malignant potential with higher drug resistance to chemodrug, Paclitaxel (PTX) and migration ability. iOVCAR-3-OSKM was maintainable and expandable on feeder-dependent culture condition, it also preserved ovarian lineage differentiation abilities, which could well differentiate into OC cells with CK-7 and CA125 expressions and develop into an organoid mimic poor prognostic OC histological feature. Conclusions The establishment of iOVCAR-3-OSKM not only allows us to fill the gap in the information on induced CICs in OC but also provides a potential strategy to develop personalized CICs and organoid models for treating OC in the near future.https://doi.org/10.1186/s12885-022-10042-3Ovarian cancer (OC)Cancer-initiating cells (CICs)Induced ovarian cancer-initiating cells (iOCICs)Organoid and drug screening |
| spellingShingle | Yu-An Chen Chen-Yu Lu Wen-Fang Cheng Kuan-Ting Kuo Chen-Wei Yu Hon-Nerng Ho Hsin-Fu Chen Szu-Hua Pan An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids BMC Cancer Ovarian cancer (OC) Cancer-initiating cells (CICs) Induced ovarian cancer-initiating cells (iOCICs) Organoid and drug screening |
| title | An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| title_full | An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| title_fullStr | An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| title_full_unstemmed | An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| title_short | An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| title_sort | experimental model for ovarian cancer propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids |
| topic | Ovarian cancer (OC) Cancer-initiating cells (CICs) Induced ovarian cancer-initiating cells (iOCICs) Organoid and drug screening |
| url | https://doi.org/10.1186/s12885-022-10042-3 |
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