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

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. Because the CAF subpopulations are likely depending on the cancer cell subtypes and aggressiveness, it is conceivable the possibility that high-metastatic cancer cells have the capacity to induce the appropriate CAF subpopulation compared with low-metastatic cancer cells. To make it clear, by using the metastatic cancer cell line models of diffuse-type gastric cancer (DGC), we cultured the immortalized stomach fibroblasts with high-metastatic and low-metastatic DGC cell lines. And then, we compared the transcriptome profile of them by microarray analysis. It has shown that high-metastatic DGC cells can induce at least two types of CAF-like phenotypes: myofibroblastic feature and chemokine producing feature. We also found that these two features of fibroblasts were basically distributed in the different fibroblasts. These data indicated that high-metastatic DGC cells form the distinct CAF subpopulations. We used two DGC cell lines: high-metastatic (44As3) and low-metastatic (HSC-44PE). The stomach fibroblasts (NF-58 and NF-60) were derived from the “normal” non-neoplastic primary tumor site of DGC patient tissue samples. These “primary” stomach fibroblasts were immortalized by following infection with retroviruses expressing mutant Cdk4, cyclin D1 and human telomerase reverse transcriptase. These immortalized fibroblasts were cultured with 44As3 or HSC-44PE. Mono-cultured fibroblasts were also prepared as a control. After one week incubation, total RNAs were extracted from fibroblasts. We used Agilent microarray chip (SurePrint G3 Human GE v3).