Thalidomide Enhances the Anti-Tumor Activity of Standard Chemotherapy in a Human Melanoma Xenotransplatation Model Elisabeth Heere-Ress, Johannes Boehm,

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Thalidomide Enhances the Anti-Tumor Activity of Standard Chemotherapy in a Human Melanoma Xenotransplatation Model Elisabeth Heere-Ress, Johannes Boehm, Christiane Thallinger, Christoph Hoeller, Volker Wacheck, Peter Birner, Klaus Wolff, Hubert Pehamberger, Burkhard Jansen Journal of Investigative Dermatology Volume 125, Issue 2, Pages 201-206 (August 2005) DOI: 10.1111/j.0022-202X.2005.23830.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Inhibition of human melanoma growth in svere combined immunodeficiency (SCID) mice by the combination of thalidomide and dacarbazine (DTIC). SCID mice were injected subcutaneously with 2.5 × 107 human melanoma cells (518 A2) into the lower left flank (n=10 animals per group). Starting one day after cell inoculation, thalidomide (administered daily at a dose of 400 mg per kg per day) or carrier solution (control) was administered for 22 d via the intraperitoneal route. From day 12 to 16, two of four groups also received daily dacarbazine (80 mg per kg per day i.p.). On day 22, the experiment was terminated. Journal of Investigative Dermatology 2005 125, 201-206DOI: (10.1111/j.0022-202X.2005.23830.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Decrease of tumor necrosis factor-α (TNF-α) level in mouse serum by thalidomide. Peripheral blood serum levels of TNF-α were measured after 22 d of treatment (carrier solution (control); thalidomide; dacarbazine (DTIC); thalidomide+DTIC). Mean TNF-α level±SD, n=10. Journal of Investigative Dermatology 2005 125, 201-206DOI: (10.1111/j.0022-202X.2005.23830.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 (a) Reduction of microvessel density by thalidomide. (a) Immunohistochemical staining of microvessels in tumor sections with CD31. A, carrier solution; B, thalidomide; C, dacarbazine (DTIC); D, thalidomide/DTIC. Figure 2 shows a representative example of tumor sections stained with CD31. The number of stained discrete microvessel in field was counted at × 200 magnification. (b) Significant reduction of microvessel in melanoma tumors by thalidomide. Statistical evaluation of the effect of thalidomide compared with carrier solution (control) on the number of microvessels in melanoma tumors grown in severe combined immunodeficiency mice. The mean number of microvessels per microscope field (0.74 mm2) was significantly reduced in the tumors treated with thalidomide and thalidomide combined with DTIC compared with the control group and the DTIC-only group. Journal of Investigative Dermatology 2005 125, 201-206DOI: (10.1111/j.0022-202X.2005.23830.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Induction of apoptosis by the combination of thalidomide and dacarbazine (DTIC). TUNEL staining of melanoma tumors grown in severe combined immunodeficiency (SCID) mice (apoptotic cells appear fluorescent green; counterstain is 4′-6′-diamidino-2-phenyllin-dole). Representative photographs of 518A2 tumors treated with (a), carrier solution (b) thalidomide, (c) DTIC, (d) thalidomide/DTIC, and (e) negative control for 22 d. Original magnification × 200. (f) Higher magnification (× 1000) to confirm apoptotic morphology. Journal of Investigative Dermatology 2005 125, 201-206DOI: (10.1111/j.0022-202X.2005.23830.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions