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CirRNA-seq of the fourth passage xenografts treated with 5-Fu and PBS

ABSTRACT: In this study, we established a successive xenograft model to enrich Epstein-Barr virus-associated gastric carcinoma (EBVaGC) cancer stem cells (CSCs) through long-term treatment of EBVaGC cell line SNU719 with 5-Fluorouraci (5-Fu) in mice vivo passage. Simply, NOD/SCID mice injected in the subcutaneous with SNU719 were treated with 5-Fu weekly until xenografts reached 1.5 cm diameter and the mice were euthanizedand single-cell suspensions were obtainedby collagenasedigestion. Then the purified cells were passaged in 5-Fu-treated NOD/SCID mice as above. After four successive generations,we obtained theSNU-4th cells from the fourth passage xenografts treated with 5-Fu. Compared with parental SNU719 cells, SNU-4th cells possessed the stemness characters. Increasing evidence have proved that circular RNA (circRNA) play an important role in maintaining the tumor phenotype. So, we compared circRNAs expression between the fourth passage xenografts treated with 5-Fu and PBS by RNA-sequencing, hoping to find out which circRNAs were involved in the stemness regulation of EBVaGC. we established a highly malignant EBVaGC cell line SNU-4th through long-term treatment of EBVaGC cell line SNU719 with 5-Fluorouraci in vivo passage, then we compared circRNAs expression between the fourth passage xenografts treated with 5-Fu and PBS by RNA-sequencing

ORGANISM(S): Homo sapiens

PROVIDER: GSE145894 | GEO | 2020/02/26

REPOSITORIES: GEO

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Project description:Fibroblast growth factor-2 delays the loss of chondrogenic potential in adult bone marrow-derived mesenchymal stem cells; We compared human mesenchymal stem cells (hMSCs), expanded long-term with and without fibroblast growth factor (FGF) supplementation, with respect to their proliferation rate, and ability to differentiate along the chondrogenic pathway in vitro. hMSCs expanded in FGF-supplemented medium proliferated more rapidly than those expanded under control conditions. Aggregates of FGF-treated cells exhibited chondrogenic differentiation at passages 1 through 7, although, in some of the preparations, chondrogenic differentiation was somewhat diminished after seventh passage. Aggregates made with control cells differentiated along the chondrogenic lineage at first passage but exhibited only marginal chondrogenic differentiation after four passages and failed to form cartilage after seven passages. Microarray analysis of gene expression identified 334 transcripts that were differentially expressed in fourth passage control cells which had reduced chondrogenic potential compared to fourth passage FGF-treated cells which retained this differentiation capacity and 243 transcripts that were differentially expressed when comparing them to first passage control cells which were also capable of differentiating into chondrocytes. The intersection of these analyses yielded 49 transcripts that were differentially expressed in cells that exhibited chondrogenic differentiation in vitro compared to cells that did not. These preliminary data must now be validated to verify whether the different gene expression profiles translate into functional differences. These findings suggest that care should be exercised when extensively expanding these cells for cartilage tissue engineering applications. Experiment Overall Design: hMSCs from three different donors were expanded for up to seven passages with or without FGF supplementation. At the end of first, fourth and seventh passages the chondrogenic potential and gene expression progfiles were analyzed.

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CirRNA-seq of the fourth passage xenografts treated with 5-Fu and PBS

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