[Colorectal cancer cells induce the formation of cancer-associated fibroblasts by activating the ERK signaling pathway in fibroblasts].

OBJECTIVE

To investigate the mechanism by which conditioned medium of colorectal cancer cells promotes the formation of cancer-associated fibroblasts (CAFs).

METHODS

Normal human colorectal fibroblasts (CCD-18Co cells) in logarithmic growth phase were treated with the conditioned media of colorectal cancer HCT116 cells (HCT116-CM) or Caco-2 cells (Caco-2-CM) alone or in combination with 300 nmol/L ERK inhibitor SCH772984. The expression levels of CAFs-related molecular markers were detected in the treated cells with real-time quantitative PCR (RT- qPCR) and immunofluorescence assay, and the changes in cell proliferation, colony formation and migration were assessed with RTCA, colony formation and wound healing assays; Western blotting was performed to detect the activated signaling pathways in the fibroblasts and the changes in CAFs formation after blocking of the signaling pathway.

RESULTS

HCT116-CM and Caco-2-CM significantly upregulated mRNA expression levels of CAFs markers (including -SMA, FAP, FN and TGF-β) in CCD-18Co cells, and strongly promoted fibroblast transformation into CAFs ( < 0.05). The two conditioned media also promoted the proliferation, colony formation and migration of CCD-18Co cells ( < 0.05) and significantly increased the levels of -SMA protein and ERK phosphorylation in the cells ( < 0.05). The ERK inhibitor SCH772984 obviously inhibited the expression of -SMA and the transformation of CCD-18Co cells into CAFs induced by the conditioned medium of colorectal cancer cells ( < 0.05).

CONCLUSION

Colorectal cancer cells may induce the formation of colorectal CAFs by activating the ERK pathway in the fibroblasts.