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Quantitative analysis of transcriptomes in irradiated/unirradiated pancreatic cancer cells

ABSTRACT: Irradiated dying tumor cells play crucial roles in tumor repopulation. However, the transcriptome changes in pancreatic cancer cells after radiation remains unknown. Here we performed RNA-sequencing in pancreatic cancer cell line SW1990 and PANC-1 that were 10Gy irradiated or unirradiated. Bioinformatic analysis revealed that the transcriptomes were significantly changed after radiation. Gene Ontology (GO) analysis revealed that the signaling concerning extracellular vesicles, especially exosomes, were significantly enriched. Further investigation revealed the vital role of irradiated dying tumor cells-derived exosomes in potentiating tumor repopulation.

ORGANISM(S): Homo sapiens

PROVIDER: GSE138984 | GEO | 2020/06/14

REPOSITORIES: GEO

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Project description:The precise mechanism of intercellular communication among cancer cells after radiation exposure remains unclear. Exosomes are membrane-enclosed small vesicles constituted by lipid bilayers and are recognized as mediators of intercellular communication that transport a variety of intracellular components, including microRNA (miRNA). Here we identified novel roles of exosomes released from irradiated cells to neighboring cancer cells. To confirm the presence of exosomes in the culture media of the human pancreatic cancer cell line MIAPaCa-2, ultracentrifugation was performed followed by transmission electron microscopy and nanoparticle tracking analysis (NanoSight) using the exosome-specific surface markers CD9 and CD63. Subsequent endocytosis of exosomes was confirmed by fluorescent microscopy. Cell survival after irradiation and following the addition of exosomes was evaluated by a colony-forming assay. Expression of miRNAs in exosomes was then quantified by microarray analysis, while those of Cu/Zn superoxide dismutase (SOD1) and Mn-superoxide dismutase (SOD2) enzymes in MIAPaCa-2 cells were evaluated by western blotting. Results showed that the uptake of irradiated exosomes was significantly higher than that of non-irradiated exosomes. Notably, irradiated exosomes induced higher intracellular levels of ROS and a higher frequency of DNA damage in MIAPaCa-2 cells, as determined by fluorescent microscopy and immunocytochemistry, respectively. Moreover, six upregulated and five downregulated miRNAs were identified in 5 Gy- and 8 Gy-irradiated cells using miRNA microarray analyses. Further analyses using miRNA-mimics and real time reverse transcription PCR identified miR-6823-5p as a possible candidate for SOD1 inhibition, leading to increased intracellular ROS level and DNA damage. This is the first study to demonstrate that irradiated exosomes can enhance the radiation effect via increased intracellular ROS levels in cancer cells, potentially leading to improved understanding of the bystander effect of neighboring cancer cells.

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Quantitative analysis of transcriptomes in irradiated/unirradiated pancreatic cancer cells

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