Christiane Thallinger, Markus F

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Mcl-1 Antisense Therapy Chemosensitizes Human Melanoma in a SCID Mouse Xenotransplantation Model Christiane Thallinger, Markus F. Wolschek, Volker Wacheck, Helmut Maierhofer, Patrick Günsberg, Peter Polterauer, Hubert Pehamberger, Brett P. Monia, Edgar Selzer, Klaus Wolff, Burkhard Jansen Journal of Investigative Dermatology Volume 120, Issue 6, Pages 1081-1086 (June 2003) DOI: 10.1046/j.1523-1747.2003.12252.x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 In vitro oligonucleotide uptake by human melanoma cells. 518A2 cells were incubated with fluorescein-labeled Mcl-1 ASO (a) 250 nM, in the presence of uptake-enhancing lipids (lipofectin); (b) 1 μM, without uptake-enhancing lipids. Twenty-four hours later, cells were fixed with paraformaldehyde and evaluated under a fluorescence microscope. Original magnification: 400×. Journal of Investigative Dermatology 2003 120, 1081-1086DOI: (10.1046/j.1523-1747.2003.12252.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 In vitro Mcl-1 downregulation in human melanoma cells. 518A2 and Mel Juso cells were incubated for 4 h with the indicated oligonucleotides or vehicle control (AS, antisense; UC, universal control, SAL, saline control) in the presence of lipofectin. Twenty-four hours later, protein was extracted and analyzed for Mcl-1 and β-tubulin expression by Western blotting. Representative data from one of three independent experiments are shown. Journal of Investigative Dermatology 2003 120, 1081-1086DOI: (10.1046/j.1523-1747.2003.12252.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Downregulation of Mcl-1 expression in human melanomas grown in SCID mice. Immunohistochemical detection of Mcl-1 in tumor xenotransplants of mice with established 518A2 tumors were treated for 14 d with the indicated oligonucleotide (a, SAL; b, UC; c, AS) delivered by miniosmotic pumps. Journal of Investigative Dermatology 2003 120, 1081-1086DOI: (10.1046/j.1523-1747.2003.12252.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Mcl-1 ASO chemosensitizes human melanoma in vivo. SCID mice with established 518A2 tumors received a 14 d infusion by miniosmotic pumps of Mcl-1 ASO (AS), universal control oligonucleotide (UC), or saline (n=6 animals per group). In addition, animals received DTIC treatment on days 12–16. (a) During the course of the experiment tumor size was assessed by caliper measurement and is given in mL as mean±SD. (b) At the end of the experiment (day 21) tumors were resected and weighed. Tumor weight is given in mg as mean±SD. Furthermore, melanoma xenografts were analyzed by TUNEL assay (apoptotic cells appear green) and counterstained with DAPI. Representative photographs of tumors are shown. Original magnification: 200×. Journal of Investigative Dermatology 2003 120, 1081-1086DOI: (10.1046/j.1523-1747.2003.12252.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions