Regulation of intestinal myofibroblasts by KRas-mutated colorectal cancer cells through heparin-binding epidermal growth factor-like growth factor

Oncol Rep. 2017 May;37(5):3128-3136. doi: 10.3892/or.2017.5520. Epub 2017 Mar 22.

Abstract

In colorectal cancer, gain-of-function mutations in KRas play a critical role in malignant transformation. Tumor growth in colorectal cancer is known to be promoted by the intestinal myofibroblasts (IMFs) that localize adjacent to the cancer cells, but the mechanisms of interaction between KRas-mutated cancer cells and the myofibroblasts remain unclear. Here, we investigated the effects of KRas-mutated cells on the behavior of myofibroblasts by using mouse primary IMFs and cells of an IMF cell line (LmcMF) and a mouse colon epithelial cell line (aMoC1). Conditioned medium (CM) was collected from aMoC1 cells overexpressing a control vector or KRasV12 vector (KRasV12-CM), and the effects of KRasV12-CM on IMFs were analyzed by performing proliferation assays, wound-healing assays, Boyden chamber assays, and western blotting. Whereas KRasV12-CM exerted little effect on the differentiation and proliferation of primary IMFs, the CM promoted migration of both primary IMFs and LmcMF cells. In KRasV12-overexpressing aMoC1 cells, mRNA expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) was higher than in mock-transfected aMoC1 cells, and HB-EGF promoted the migration of primary IMFs and LmcMF cells. Moreover, KRasV12-CM-induced IMF migration was suppressed by dacomitinib, an inhibitor of HB-EGF receptors. Notably, in LmcMF cells, both KRasV12-CM and HB-EGF activated extracellular signal-regulated kinase (ERK) and c-jun N-terminal kinase (JNK), whereas KRasV12-CM-induced migration of IMFs was suppressed following treatment with either an ERK inhibitor (FR180204) or a JNK inhibitor (SP600125). These results suggest that HB-EGF secreted from KRas-mutated colorectal cancer cells promotes IMF migration through ERK and JNK activation, which, in turn, could support cancer progression.

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Movement
  • Cell Proliferation / drug effects
  • Colorectal Neoplasms / genetics*
  • Culture Media, Conditioned / pharmacology
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Heparin-binding EGF-like Growth Factor / metabolism*
  • Humans
  • Intestinal Mucosa / metabolism
  • Intestines / cytology*
  • MAP Kinase Kinase 4 / metabolism
  • Mice
  • Mutation*
  • Myofibroblasts / cytology*
  • Myofibroblasts / drug effects
  • Proto-Oncogene Proteins p21(ras) / genetics*

Substances

  • Culture Media, Conditioned
  • Hbegf protein, mouse
  • Heparin-binding EGF-like Growth Factor
  • Extracellular Signal-Regulated MAP Kinases
  • MAP Kinase Kinase 4
  • Hras protein, mouse
  • Proto-Oncogene Proteins p21(ras)