Journal: Cancer gene therapy

Article Title: Blood outgrowth endothelial cell-based systemic delivery of antiangiogenic gene therapy for solid tumors

doi: 10.1038/cgt.2010.42

Figure Lengend Snippet: Inhibition of endothelial cell proliferation in vitro. A proliferation assay was performed on PVECs exposed to conditioned medium from transduced BOECs expressing angiostatin and endostatin (AE), mouse soluble Flt-1 receptor (msFlt1) or DsRed or a medium from a mixed BOEC culture (AE+msFlt1). Results are relative to PVECs cultured with basal medium supplemented with CAMP and FGF. Data points represent the means ± S.E. of six wells from one experiment.

Article Snippet: Serum samples were collected at the time of animal sacrifice, and protein levels were measured with a murine endostatin ELISA kit (Chemicon International).

Techniques: Inhibition, In Vitro, Proliferation Assay, Expressing, Cell Culture

Journal: The Journal of Immunology Author Choice

Article Title: Inflammation Induced by MMP-9 Enhances Tumor Regression of Experimental Breast Cancer

doi: 10.4049/jimmunol.1202610

Figure Lengend Snippet: Dose-dependent effects of MMP-9 gene transfer. Nude mice were oophorectomized and supplemented with a physiologic level of estradiol. MCF-7 cells were injected s.c., and tumors were formed on the right hind flank. At similar tumor sizes (∼40 mm2), gene transfer by adenovirus was performed at different concentrations, PFU, with Addl70-3 (empty control vector) or AdMMP-9 vector. (A) Dose-dependent effects on tumor growth and microvessel area by AdMMP-9. AdMMP-9 decreased tumor growth in a dose-dependent manner; ***p < 0.0001; AdMMP-9 0.5 × 109 PFU and AdMMP-9 2 × 109 PFU versus control; n = 6–12, ANOVA, Bonferroni post hoc test. #p < 0.05, AdMMP-9 0.5 × 109 PFU versus AdMMP-9 2 × 109 PFU; n = 6–8, t test. Quantification of microvessel area using staining for von Willebrand factor revealed decreased microvessel area between the treatment groups. n = 15, ANOVA, Bonferroni post hoc test. AdMMP-9 versus control, ***p < 0.0001, n = 15 and AdMMP-9 0.5 × 109 PFU versus AdMMP-9 2 × 109 PFU, ###p < 0.0001. (B) In vivo microdialysis sampling of VEGF and endostatin after AdMMP-9. No significant increases of stroma-derived (murine) VEGF and cancer cell–derived (human) VEGF were found. ANOVA, n = 4–8 in each group. Significant increased levels of both murine and human endostatin: *p < 0.05, ***p < 0.0001 compared with control. (C) Dose-dependent increase of neutrophil infiltration by AdMMP-9. Top panel, After immunohistochemistry, as described in the Materials and Methods section, the number of neutrophils was counted on tumor sections and shown to increase significantly after AdMMP-9 gene transfer in a dose-dependent fashion. Representative sections from each treatment group are depicted. n = 12, ANOVA, Bonferroni post hoc test. *p < 0.05, ***p < 0.001 compared with control, #p < 0.05 compared with 0.5 × 109 PFU AdMMP-9. Bottom panel, The chemokines KC and MIP-2 increased after AdMMP-9 treatment: n = 6–8 in each group, AVOVA, Bonferroni post hoc test. Scale bars, 20 μm. *p < 0.05, **p < 0.01, ***p < 0.001 compared with control. (D) In vivo cytokine profile by AdMMP-9. One week after gene transfer, microdialysis was performed to sample proteins in live tumor tissue in vivo. An M1-like cytokine profile was detected after AdMMP treatment. n = 6–8 in each group, ANOVA, Bonferroni post hoc test. *p < 0.05, **p < 0.01, ***p < 0.0001 compared with control.

Article Snippet: Quantification of proteins in microdialysates and cell-culture media Microdialysates and culture media were analyzed with immunoassays from R&D Systems (Minneapolis, MN) unless otherwise stated: human endostatin (Quantikine), murine endostatin (murine ELISA kit; Uscn life Science, Wuhan, China), human vascular endothelial growth factor (VEGF; QuantiGlo), and murine VEGF (mouse VEGF; Quantikine).

Techniques: Injection, Plasmid Preparation, Staining, In Vivo, Sampling, Derivative Assay, Immunohistochemistry