Journal Article

An experimental model for ovarian cancer: propagation of ovarian cancer initiating cells and generation of ovarian cancer organoids

Kuo, Kuan-Ting, Ho, Hon-Nerng, Cheng, Wen-Fang, Yu, Chen-Wei, Chen, Yu-An, Lu, Chen-Yu, Pan, Szu-Hua, Chen, Hsin-Fu

2022

Overview

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.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Animal research / Biomedical and Life Sciences / Biomedicine / Cancer cells / Cancer Research / Cancer therapies / Cancer-initiating cells (CICs) / Cell culture / Cell differentiation / Chemotherapy / Development and progression / Diagnosis / Drug development / Drug resistance / Gene expression / Growth / Gynecological cancer / Gynecology / Health Promotion and Disease Prevention / Immunohistochemistry / Induced ovarian cancer-initiating cells (iOCICs) / KLF4 protein / Medical prognosis / Medical research / Medicine/Public Health / Metastasis / Myc protein / Oct-4 protein / Oncology / Organoid and drug screening / Organoids / Ovarian cancer / Ovarian cancer (OC) / Paclitaxel / Patients / Polymerase chain reaction / Propagation / Stem cells / Surgery / Surgical Oncology / Transcription factors / Tumorigenesis / Tumors