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Figure 4 | Cancer Cell International

Figure 4

From: Paracrine factors from adipose-mesenchymal stem cells enhance metastatic capacity through Wnt signaling pathway in a colon cancer cell co-culture model

Figure 4

The Wnt signaling plays a key role in the production of paracrine factors of AMSCs malignancy of colon cancer cells in co-culture model. HCT116 cells were co-cultured with AMSCs in the presence of Wnt signaling inhibitor IWP2, and the production of paracrine factors of AMSCs and the malignancy of HCT116 cells were investigated in vitro and/or in vivo assays. (A) Representative immunoblots of nuclear beta-catenin from HCT116 cells cultured in indicated conditions (top panel) and their fold changes quantified by a densitometry analysis (bottom panel). (B-C) The alterations of transcripts of indicated EMT- and stemness-related genes of HCT116 cells (B), and indicated cytokines and MMPs genes of AMSCs in different conditions (C). (D) The concentration of indicated paracrine factors in the culture medium. (E) The capacity of invasion of HCT116 cells in the indicated conditions. Left panel: representative images of indicated condition; right panel: numbers of invaded cells in indicated conditions. (F) The capacity of colony formation of HCT116 cells in the indicated. Left panel: representative images of indicated condition; right panel: numbers of sphered cells in indicated conditions. (G) The in vivo tumorigenitic assay showed a repression of tumorigenicity of HCT116 cells in co-culture model with IWP2 as ascertained by the size (left panel) and weight (right panel) of subcutaneous primary tumors derived from SCID mice at 8 weeks after the injection of HCT116 cells. (H) Representative H&E and IHC staining of E-cadherin and vimentin in the primary tumors derived from SCID mice in (G). Scale bars represent 20 μm. GAPDH was used as an endogenous control for normalization. Data represented as mean ± SD from three independent triplicated experiments (N = 9). Compared to the control, *p < 0.05; **p < 0.01; ***p < 0.001.

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