Fig. 10. Suppression of both tumor growth and tumor-induced angiogenesis by transduction of Ad-shChM-I or Ad-shTeM. (A) C57BL/6 mice at 21 days following implantation of melanoma cells (BL-6). Red circles indicate implanted tumors. Scale bar: 1.2 cm. (B) Images of subcutaneously injected melanoma tumor cells, transduced with or without Ad-shChM-I, Ad-shTeM and Ad-EGFP, were photographed also on day 21. (C) Time course of increase in tumor weight following implantation. At 7, 14 and 21 days after subcutaneous implantation of both transduced and non-transduced melanoma cells, the tumor volume was determined in Ad-shChM-I (open boxes), Ad-shTeM (open circles)-, Ad-EGFP (filled boxes)-transfected groups or in the wild-type group. The plots represent the mean values of tumor volumes from four animals in each group. Bars indicate±s.d. (D) Reduced angiogenesis in Ad-shChM-I or Ad-shTeM transduced tumors is apparent. Vasculariza tion in tumors on day 21 following implantation is visualized by staining of PECAM-1 with monoclonal antibodies. Cell nuclei are stained blue-violet with Hoechst dye 33342 as. Scale bar: 400 µm. (E) Microvessel numbers were counted from four different random fields of both adenovirus transduced or non-transduced melanoma cells. Data are represented as mean values (±s.d.) of microvascular densities per field (x40) from sections of Ad-shChM-I-, Ad-shTeM-, Ad-EGFP-transduced or non-transduced tumors (P<0.005).