Normal human colonic epithelial cells NCM‐460 and CC cell lines including SW620, HT29, LoVo and SW1116 were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA). All cells were grown in Ham's F12 medium (GIBCO, Barcelona, Spain) which was supplemented with 7% foetal bovine serum (FBS) under humidified environment at the controlled temperature of 37°C and atmosphere of 5% CO2.

The CC tissues were minced and re‐suspended in a Dulbecco's modified eagle's medium (DMEM) containing penicillin‐streptomycin (Invitrogen, Carlsbad, CA, USA), amphotericin B (Invitrogen, Carlsbad, CA, USA), 3% collagenase and 20% FBS (Gibco, Grand Island, NY, USA) for 2 hours. The sample was filtered using an 8‐μm filter for 2 hours for the removal of undigested debris. Single‐cell suspension that was supplemented with viable fibroblasts was plated in 24‐well plates in DMEM containing 10% FBS for duration of 2‐3 weeks and then transferred to T75 flasks for further culture. The tenth passage of fibroblasts was used for subsequent experiments. Normal fibroblasts (NFs) were obtained 10 cm from the edge of the tumour infiltration. The morphology of isolated CAFs and NFs was observed under an inverted microscope. CAFs and NFs were identified through the determination of expression of α‐smooth muscle actin (α‐SMA), fibroblast activation protein (FAP), fibroblast‐specific protein 1 (FSP‐1) and vimentin using immunofluorescence and Western blot analysis.

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