Topically Applied Imiquimod Inhibits Vascular Tumor Growth In Vivo

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Topically Applied Imiquimod Inhibits Vascular Tumor Growth In Vivo Robert Sidbury, Nicole Neuschler, Erin Neuschler, Ping Sun, Xiao-qi Wang, Elena Puscasiu, Sajiv Gugneja Journal of Investigative Dermatology Volume 121, Issue 5, Pages 1205-1209 (November 2003) DOI: 10.1046/j.1523-1747.2003.12521.x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Imiquimod inhibits tumor growth. Mice were treated with imiquimod cream three times weekly beginning on day 1 (I1) or day 5 (I5) after tumor cell inoculation, vehicle base beginning on day 1 (B1) or day 5 (B5) after inoculation, or with 20% arachidonic acid beginning day 1 (A1) after inoculation. Tumors were measured every other day once visible and tumor volumes calculated. Error bars represent mean±SD. I, imiquimod; B, vehicle base; A, arachidonic acid. Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Topical application of imiquimod inhibits tumor growth in 129/J mice. Tumors were photographed serially every 4 d to record the appearance of the tumors and the potential irritant dermatitis. Mice treated with imiquimod beginning day 1 at 9 d (a) and 17 d (c) after EOMA cell introduction. Mice treated with vehicle base beginning on day 1 at 9 d (b) and 17 d (d) after EOMA cell introduction. Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Imiquimod application to mouse tumors increases mouse survival. Imiquimod, or as a control the vehicle base or arachidonic acid, was applied three times weekly beginning on day 1 to tumors. Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Imiquimod treatment inhibits tumor PCNA expression. PCNA expression was analyzed by immunohistochemistry using anti-mouse PCNA monoclonal antibody. I1-I4 are samples extracted from four EOMA cell tumors treated with imiquimod beginning on day 1; B1–4 are samples extracted from four tumors treated with vehicle base. Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Imiquimod application triggers vascular tumor cell apoptosis. Tumors from mice treated with imiquimod (a) or vehicle base (b) were evaluated for apoptotic cells by immunohistochemical staining with a Klenow FragEL DNA fragmentation detection kit. Bar=50 μm. Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Imiquimod treatment decreases MMP-9 activity through enhancement of TIMP-1 expression. Tumor tissue of mice treated with imiquimod (I) or vehicle base (B) was either treated with lysis buffer to extract total protein for TIMP-1 and MMP-9, or incubated in serum-free Dulbecco's modified essential medium to analyze MMP-9 and MMP-2 activities as described in Materials and Methods. TIMP-1 expression was detected using anti-mouse TIMP-1 monoclonal antibody (a). MMP-9 expression was detected with anti-mouse MMP-9 antibody (b, top row). MMP-9 and MMP-2 activities were determined by zymography using gelatin as a substrate (b, middle and bottom rows, respectively). Journal of Investigative Dermatology 2003 121, 1205-1209DOI: (10.1046/j.1523-1747.2003.12521.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions