Fig 1: CD133+ HPCs affected the expression profiles of cytokines/chemokines/MMPs by cancer cells. HPCs were co-cultured with MDA-MB-435s-HM or MDA-MB-435s cells. The concentrations of CXCL16, IL-2Rα, IL-2Rγ, MMP-1, MMP-9, PDGFR-α, SDF-1α, TGF-β, PECAM-1, and VE-cadherin in the supernatant of MDA-MB-435s-HM cells were significantly higher than those in the supernatant of MDA-MB-435s cells (> fivefold, a–j). The concentrations of MMP-9, PDGFR-α, and PECAM-1 in the supernatant of MDA-MB-435s-HM cells were significantly elevated after co-culture of MDA-MB-435s-HM cells with CD133+ HPCs cells (e, f, i). The concentrations of CXCL-16, IL-2Rα, MMP-1, MMP-9, PDGFR-α, and PECAM-1 in the supernatant of CD133+ HPCs were elevated when MDA-MB-435s-HM cells were co-cultured with CD133+ HPCs (a, b, d, f, i)
Fig 2: Effect of the inhibition of MMP-9, PDGFR-α, and PECAM-1 on the behaviors of MDA-MB-435s-HM cells. HPCs were co-cultured with MDA-MB-435s-HM cells. a Representative images of MDA-MB-435s-HM cell migration. TIMP-1, PDGFR tyrosine kinase inhibitor III, and WM59 were added to the culture medium in the experiment group, and normal saline was added to the control group. A significantly decreased number of migrated cells could be seen in the experimental group. b Statistical analysis of cell migration. c Inhibition of MMP-9, PDGFR-α, and PECAM-1 suppresses lung metastasis of MDA-MB-435s-HM cells
Fig 3: Validation of protein microarray results by ELISA and qRT-PCR. HPCs were co-cultured with MDA-MB-435s-HM or MDA-MB-435s cells. a The ELISA results showed trends consistent with those observed by protein microarray in the concentrations of IL-2Rα, MMP-1, MMP-9, PDGFR-α, TGF-β, PECAM-1, and VE-cadherin among the four groups. However, there were no significant differences in CXCL-16, IL-2Rγ and SDF-1α expression among the four groups. b The trends in the mRNA expression of IL-2Rα, MMP-1, MMP-9, PDGFR-α, TGF-β, PECAM-1, and VE-cadherin among the four groups were completely consistent with protein concentration results obtained by ELISA
Fig 4: Inhibition of MMP-9, PDGFR-α, and PECAM-1 decreases the amount of VEGFR1+CD133+ HPCs in the lung. The peak of the average ratio of VEGFR1+ HPCs was postponed and synchronous to the appearance of detectable lung metastasis. Transfection of miR-140-3p enhanced the antitumor effect of sorafenib on MHCC97-H cells’ intrahepatic growth
Fig 5: HPCs enhance the invasive growth of MDA-MB-435s-HM cells. HPCs were co-cultured with MDA-MB-435s-HM cells, and cells were treated with an inhibitor of MMP-9, anti-CD31 antibody, or an inhibitor of PDGFR-α. The invasive growth was assessed based on aMicroPET photographs, area of nodules in liver formed by MDA-MB-435s-HM cells or Masson staining indicating the invasive growth of cells. Quantitative results from bMicroPET photographs; c fold difference in radioactivity between liver and blood; d inhibitory rates of inhibitors on 18F-FDG intensity or e invasive growth of cells. Definitions: inhibitor of MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) shown to be a natural inhibitor of MMP-9; anti-CD31 antibody, anti-human CD31 antibody recognizing the D2 extracellular portion of CD31 to block its function; inhibitor for PDGFR-α, PDGFR tyrosine kinase inhibitor III shown to be an ATP-competitive inhibitor of PDGFR-α. *P < 0.05. White arrow in Figure indicates the nodules in mouse liver
Supplier Page from Abcam for Human CD31 ELISA Kit (PECAM)