Abstract:
Objective To investigate whether the omental-adipose stromal cells (O-ASCs) exposing to gastric cancer-conditioned medium (CM) could be inducted to differentiate into carcinoma-associated fibroblasts (CAFs) and the effect of ERK signaling pathway in the process.
Methods We identified O-ASCs by examining their ability to differentiate osteogenic and adipogenic lineages and through flow cytometry. O-ASCs were co-cultured with MGC803 and SGC7901CM. The expression of CAFs markers (α-SMA, FSP-1, and vimentin) and paracrine factors (VEGFA, TGF-β, FAP, and SDF-1) were evaluated by RT-PCR and Western blot. In vitro cultures of OASCs were divided into three groups: the control, SGC7901-CM, and SGC7901-CM+U0126 groups. Cells were collected after 12 h. Western blot was performed to evaluate the expression of α-SMA, FSP-1, ERK, and p-ERK1/2.
Results The primary cells were O-ASCs. The expression levels of CAFs markers (α-SMA, FSP-1, and vimentin) and O-ASC paracrine factors (VEGFA, TGF-β, FAP, and SDF-1) clearly increased (P < 0.05). In comparison with the control, the expression of ERK in SGC7901-CM group did not change (P > 0.05), while the expression of p-ERK1/2, α-SMA, and FSP-1 significantly improved (P < 0.05). Comparison of SGC 7901-CM + U0126 and SGC 7901-CM groups showed that the expression levels of ERK had no statistical difference (P > 0.05), while the expression levels of p-ERK1/2, α-SMA, and FSP-1 decreased (P < 0.05).
Conclusion O-ASCs participate in the peritoneal metastasis of gastric cancer through differentiation by CAF and paracrine factors. The ERK signaling pathway is important in the differentiation of O-ASCs towards CAFs.
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