The transcriptome profile of the stomach fibroblasts with high-metastatic or low-metastatic gastric cancer cell-derived extracellular vesicles.

Cancer-associated fibroblasts (CAFs) have an important role in the tumor progression. CAFs are heterogeneous, and its subpopulation with distinct functions have been regarded as the major obstacle to CAF targeting therapy. However, it is still unclear how cancer cells mediate CAF subpopulations and create preferable tumor microenvironment for their metastasis. In this study, by using the metastatic cancer cell line models of diffuse-type gastric cancer (DGC), we demonstrate that high-metastatic cancer cell-derived extracellular vesicles (EVs) contribute to the formation of activated fibroblast subpopulations. High-metastatic DGC cells create at least two types of CAF subpopulations: myofibroblastic feature and chemokine producing feature. The CAF subpopulation with chemokine producing feature was selectively induced by high-metastatic DGC cell-derived EVs. We also found that high-metastatic DGC cell-derived EVs contain various miRNAs to induce chemokine expression in the fibroblasts. Our findings suggest that intercellular communications via EVs contribute to establishing the appropriate tumor microenvironment toward cancer metastasis. To study the effect of EVs derived from GC cell lines on gene expression in the stomach fibroblasts, transcriptome analysis for the stomach fibroblasts with EVs derived from GC cell lines was performed. EVs were isolated from the conditioned media of two DGC cell lines: high-metastatic (44As3) and low-metastatic (HSC-44PE). The stomach fibroblast was derived from the “normal” non-neoplastic primary tumor site of DGC patient tissue sample. The “primary” stomach fibroblast was immortalized by following infection with retroviruses expressing mutant Cdk4, cyclin D1 and human telomerase reverse transcriptase. DGC cell-derived EVs were treated to the immortalize stomach fibroblasts. PBS (-) treatment and no treated samples were also prepared as a control. After five days incubation, total RNAs were extracted from them. We used Agilent microarray chip (SurePrint G3 Human GE v3).