Figure 1
Adipocytes secreted factors induce EMT and aggressiveness in in vitro models of pancreatic preneoplastic lesions. (a) Isolation and adipogenic differentiation of hBMSCs. The morphology of adherent hBMSCs and adipogenic differentiation. The accumulation of cytoplasmic triglycerides was detected by Oil Red O staining and visualized under a microscope. (b) Quantitative real-time PCR analysis of leptin (Lep) and adiponectin (ADIPOQ) along adipogenic differentiation. The values represent the fold change in RNA expression between the gene of interest and b-actin. (c) Western blot analysis of EMT marker vimentin in HPDE models cultured with conditional medium (CM) from hBMSC (hBMSCCM) and adipocytes (ADIPOCM). γ-Tubulin was used as loading control. (d) Cell proliferation assay of HPDE and HPDE/KRAS cell lines cultured with the conditional medium from hMBSC and adipocytes. The mean values and 95% confidence interval (CI) from three independent experiments done in quadruplicate are shown. (e) Levels of cell migration in HPDE and HPDE/KRAS cell lines co-cultured with hBMSC and adipocytes. The mean values and 95% CI from three independent experiments done in quadruplicate are shown. ***P<0.001 by two-tailed unpaired Student’s t-tests. Photographs of the wound area were taken by using phase-contrast microscopy immediately at indicated time point after the incision. All these experiments were performed by using hBMSC isolated from the healthy donor 1 (#1) and its relative adipocytes.
