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SSEA-4 labels a subset of poor prognosis-associated osteosarcoma-initiating cells that respond to differentiation induction

ABSTRACT: Osteosarcoma is thought of arising from the malignant transformation of osteogenic progenitors. The stage-specific embryonic antigen-4 (SSEA-4) labels a subset of TICs specially present in the high-risk subgroup.

ORGANISM(S): Homo sapiens

PROVIDER: GSE58209 | GEO | 2014/06/05

SECONDARY ACCESSION(S): PRJNA251592

REPOSITORIES: GEO

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Project description:In the cancer stem cell model a cell hierarchy has been suggested as an explanation for intratumoral heterogeneity, and tumor formation is thought to be driven by this tumor cell subpopulation. The identification of cancer stem cells in osteosarcoma (OS) and the biological processes dysregulated in this cell subpopulation, also known as tumor-initiating cells (TICs), may provide new therapeutic targets. The goal of this study, therefore, was to identify and characterize the gene expression profiles of TICs isolated from human OS cell lines. We analyzed the self-renewal capacity of OS cell lines and primary OS tumors based upon their ability to form sphere-like structures (sarcospheres) under serum-starving conditions. TICs were identify from OS cell lines using the long-term label retention dye PKH26. OS TICs and the bulk of tumor cells were isolated and used to assess their ability to initiate tumor in NOD/SCID mice. Gene expression profiles of OS TICs were obtained from fresh orthotopic tumor samples. We observed that increased sarcosphere efficiency correlated with an enhanced tumorigenic potential in OS. PKH26High cells were shown to constitute OS TICs based upon their capacity to form more sarcospheres, as well as to generate both primary bone tumors and lung metastases efficiently in NOD/SCID mice. Genomic profiling of OS TICs revealed that both bone development and cell migration processes were dysregulated in this tumor cell subpopulation. PKH26 labeling represents a valuable tool to identify OS TICs and gene expression analysis of this tumor cell compartment evidences potential therapeutic targets. Gene expression analysis was performed in PKH26Hi cells (TICs) and PKH26Lo cells (non-TICs) obtained from five orthotopic bone tumors generated by each MNNG/HOS and 143B cell lines.The soft tissue component from each bone tumor was dissected, minced and processed. Immediately after FACS, 1.5x10^5 PKH26Hi cells and 7x10^5 PKH26Lo cells from each bone tumor sample were snap frozen and stored at -80°C until RNA processing. The microarray data were first evaluated for quality control using the AffyQCReport library of BioConductor 50.Two 143B PKH26Hi tumor samples were processed together due to insufficient RNA material.

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SSEA-4 labels a subset of poor prognosis-associated osteosarcoma-initiating cells that respond to differentiation induction

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