Skip to main content
. 2009 Apr 15;101(8):592–604. doi: 10.1093/jnci/djp058

Figure 4.

Figure 4

Expression of TSP-1 and tumor growth in nude mice. A) Tumor volume and TSP-1 expression. The 2MLN-dnTβRII cells, which stably express TSP-1, are termed 2MLN-dnTβRII+TSP-1. Volumes of the subcutaneous tumors produced by 2MLN-GFP, 2MLN-dnTβRII, and 2MLN-dnTβRII+TSP-1 cells were determined 7 days after inoculation (n = 6 mice per group). B) Vascular density at 7 days after inoculation as determined by immunostaining for PECAM-1. Left) Immunostaining with antibody against PECAM-1. Scale bars = 100 μm. Right) Percent PECAM-1–positive area (n = 3 with each condition). C) Fibrotic tissue as determined by AZAN staining in the subcutaneous tumors 7 days after inoculation. Left) Micrographs with fibrotic tissue stained blue by AZAN staining. Scale bars = 100 μm. Right) Percent AZAN-positive area (n = 3 with each condition). D) The effect of miTSP-1 mRNA expression in the gastric cancer cells as determined by TSP-1 mRNA expression. The 2MLN-GFP, 2MLN-GFP+miTSP-1, and 2MLN-dnTβRII cells were treated with TGF-β or left untreated for 24 hours in vitro, and expression of TSP-1 mRNA was compared among the cell lines. The experiment was conducted two times, each sample was assayed in triplicate, and data were averaged. Data from one representative experiment are shown. E) Growth curves of 2MLN-GFP, 2MLN-GFP+miTSP-1, and 2MLN-dnTβRII xenograft tumors in nude mice (n = 6 in each condition). In 2MLN-GFP+miTSP-1 cells, the expression of TSP-1 was reduced by use of the miTSP-1. Tumor volume is shown relative to the average volume in each condition at day 0 after starting evaluation. F) Expression of TSP-1 mRNA in the 2MLN-GFP, 2MLN-GFP+miTSP-1, and 2MLN-dnTβRII tumors in vivo. Experiment was conducted two times, each sample was assessed in triplicate, and data were averaged. Data from one representative experiment are shown. Error bars = 95% confidence intervals. All P values (two-sided), except for those in (E), were calculated using the Student's t test. P values in (E) were calculated by two-way repeated measures analysis of variance. dnTβRII = dominant-negative TGF-β type II receptor; miTSP-1 = microRNA against thrombospondin-1; GFP = green fluorescent protein; TGF-β = transforming growth factor β; TSP-1 = thrombospondin-1; PECAM-1 = platelet–endothelial cell adhesion molecule-1.