Discovery of YS-1 as a cell line of gastric inflammatory cancer-associated fibroblasts

Abstract

Background: Inflammatory cancer-associated fibroblasts (iCAFs) was first identified by co-culture of pancreatic stellate cells and tumor organoids. The key feature of iCAFs is IL-6^high/αSMA^low. We examine this phenomenon in gastric cancer using two cell lines of gastric fibroblasts (HGF and YS-1).

Methods and results: HGF or YS-1 were co-cultured with MKN7 (a gastric adenocarcinoma cell line) in Matrigel. IL-6 protein levels in the culture supernatant were measured by ELISA. The increased production of IL-6 was not observed in any of the combinations. Instead, the supernatant of YS-1 exhibited the higher levels of IL-6. YS-1 showed IL-6^high/αSMA (ACTA2)^low in real-time PCR, mRNA-seq and immunohistochemistry. In mRNA-seq, iCAFs-associated genes and signaling pathways were up-regulated in YS-1. No transition to myofibroblastic phenotype was observed by monolayer culture, or the exposure to sonic hedgehog (SHH) or TGF-β. YS-1 conditioned medium induced changes of morphology and stem-ness/differentiation in NUGC-3 (a human gastric adenocarcinoma cell line) and UBE6T-15 (a human bone marrow-derived mesenchymal stem cell line).

Conclusions: YS-1 is a stable cell line of gastric iCAFs. This discovery will promote further research on iCAFs for many researchers.

Keywords: Cell line; Gastric cancer; Inflammatory cancer associated fibroblasts (iCAFs); Interleukin-6 (IL-6); Leukemia inhibitory factor (LIF).