A short-term colorectal cancer sphere culture as a relevant tool for human cancer biology investigation

Abstract

Background:Ex vivo colospheres have been previously characterised as a colorectal cancer (CRC) well-rounded multicellular model, exclusively formed by carcinoma cells, and derived from fresh CRC tissue after mechanical dissociation. The ability to form colospheres was correlated with tumour aggressiveness. Their three-dimensional conformation prompted us to further investigate their potential interest as a preclinical cancer tool.Methods:Patient-derived CRC xenografts were used to produce numerous colospheres. Mechanism of formation was elucidated by confocal microscopy. Expression analysis of a panel of 64 selected cancer-related genes by real-time qRT–PCR and hierarchical clustering allowed comparison of colospheres with parent xenografts. In vitro and in vivo assays were performed for migration and chemosensitivity studies.Results:Colospheres, formed by tissue remodelling and compaction, remained viable several weeks in floating conditions, escaping anoikis through their strong cell–cell interactions. Colospheres matched the gene expression profile of the parent xenograft tissue. Colosphere-forming cells migrated in collagen I matrix and metastasised when subrenally implanted in nude mice. Besides, the colosphere responses to 5-fluorouracil and irinotecan, two standard drugs in CRC, reproduced those of the in vivo original xenografts.Conclusion:Colospheres closely mimic biological characteristics of in vivo CRC tumours. Consequently, they would be relevant ex vivo CRC models.

X-linked inhibitor of apoptosis-associated factor l (XAFl) enhances the sensitivity of colorectal cancer cells to cisplatin

“…The advantages for cancer cells included convenience source, limitless passages, and easily clutured in a large-scale. Nonetheless, the findings of previous studies concluded from cancer cell models did not reflect the real responsive mechanisms during CRC chemotherapy [23,24]. The discrepancy between cancer cell lines and cancer tissues is partially resulted from variation during continuous passages [25].…”

Patient-Derived Xenografts from Non–Small Cell Lung Cancer Brain Metastases Are Valuable Translational Platforms for the Development of Personalized Targeted Therapy

“…Herein, PDXs derived from NSCLC brain metastases gave rise to numerous tumor spheres in a few days after mechanical dissociation ex vivo in a reproducible manner, whereas PDXs from NSCLC primary tumor demonstrated poor viability and sphereforming ability in vitro. In previous study, the ability to form tumor spheres was also correlated with tumor aggressiveness, and the chemosensitivity of tumor spheres was similar to that of the in vivo original xenografts (50). Therefore, ex vivo tumor spheres would be one of the potential preclinical tools for the validation of new drugs in a setting that more closely resembles patients' tumors but also for the investigation of patient-tailored therapeutics with high-throughput drug screen.…”

“…However, afatinib reportedly exhibits therapeutic effects on patients with NSCLC with EGFR exon-20 duplication (insertion) mutations (29), which was reproduced in vitro in this study. In addition, tivantinib, an MET-targeted agent, showed a similar IC 50 (270 nmol/L) for the proliferation of LC-MBT-15 cells to that of afatinib. Interestingly, the sensitivity was correlated with decreased phosphorylation of ERK regardless of the primary therapeutic targets (Fig.…”

“…Treatment with EGFR-targeted agents reduced the phosphorylation of EGFR; however, AKT phosphorylation, a downstream signaling component of EGFR, was decreased by only afatinib, which showed the lowest IC 50 for the proliferation of LC-MBT-19 cells (Table 3 and Fig. 3C).…”

“…3C). Although EGFR wild-type adenocarcinomas have been reported to be resistant to EGFR-targeted agents clinically, afatinib had therapeutic activities against this patient population (30, 31), which could draw the lowest IC 50 . Interestingly, the activation status of ERK was not affected by any of the EGFRtargeted agents.…”