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Comparison of a Treatment Strategy Combining CCI-779 Plus DTIC Versus DTIC Monotreatment in Human Melanoma in SCID Mice  Christiane Thallinger, Johannes.

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Presentation on theme: "Comparison of a Treatment Strategy Combining CCI-779 Plus DTIC Versus DTIC Monotreatment in Human Melanoma in SCID Mice  Christiane Thallinger, Johannes."— Presentation transcript:

1 Comparison of a Treatment Strategy Combining CCI-779 Plus DTIC Versus DTIC Monotreatment in Human Melanoma in SCID Mice  Christiane Thallinger, Johannes Werzowa, Wolfgang Poeppl, Florian M. Kovar, Barbara Pratscher, Peter Valent, Peter Quehenberger, Christian Joukhadar  Journal of Investigative Dermatology  Volume 127, Issue 10, Pages (October 2007) DOI: /sj.jid Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Expression of mTOR, Akt, PTEN, and S6K1 in human melanoma cells. (a) Relevant key parts of the mTOR/Akt pathway were evaluated by Western blot (10% polyacrylamide gel). The target structure of CCI-779, namely p-mTOR, was expressed in all three melanoma cell lines. PTEN, a tumor suppressor, was most weakly expressed in Mel-JUSO and strongest in 607B human melanoma cell lines. (b) Treatment of human melanoma cells with CCI-779 increased the expression of p-Akt, when compared to untreated controls in all three melanoma cell lines evaluated. In contrast, protein expression of p-S6K1 decreased when treated with CCI-779. Each lane was loaded with 10μg protein. Representative blots of different gels are shown. (c) Quantification of Western blot bands by densitometry. Data are presented as a percentage of controls (actin). Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Effect of CCI-779 treatment on melanoma growth in SCID mice. Depending on the cell lines used, the overall observation period varied from 23 to 34 days. SCID mice with 518A2, Mel-JUSO, or 607B melanomas received a 14-day treatment (day 1–14) with CCI-779 (1.5mg/kg intraperitoneally) or solvent control (6–10 animals per group). In addition, animals received 80mg/kg of DTIC intraperitoneally on treatment days 4–8. At the end of the experiment, mice were killed and tumors were resected and weighed (right panel). Tumor weight is provided in grams. During the course of the experiment, tumor size was assessed by caliper measurements (left panel) twice weekly and is shown in mm3 (mean±SD; =statistically significant, P<0.05; n.s.=not significant, P>0.05). Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Treatment with CCI-779 plus DTIC facilitates apoptosis in vivo. Melanoma xenografts treated with either the combination of CCI-779 plus DTIC or DTIC solely were analyzed by TUNEL assay for apoptotic cell death. TUNEL-positive cells (green) increased in tumors treated with CCI-779 plus DTIC in Mel-JUSO and 607B xenografts compared to DTIC monotreatment. (Bar=0.1mm.) Representative photographs of tumor sections are shown. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Expression of p70 S6K before and after treatment with CCI-779 in vivo. Melanoma xenografts treated with either CCI-779 or solvent control were analyzed by immunohistochemistry for cytoplasmic and nuclear p70 S6K protein expression. Cytoplasmic p70 S6K expression impressively decreased after CCI-779 treatment, whereas the nuclear expression pattern remained nearly unaffected. (Bar=0.1mm upper panel; bar=0.015mm lower panel.) Representative photographs of tumor sections are shown. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Expression of mTOR, Akt, PTEN, and S6K1 in human melanomas grown in SCID mice. p-mTOR and PTEN protein expression of human melanomas grown in SCID mice were not influenced by the treatment with CCI-779. In contrast, treatment of human melanomas with CCI-779 increased the expression of p-Akt when compared to untreated controls and induced additionally a slight decrease of p-S6K1 protein expression (15μg protein were loaded per lane). Representative blots of different gels are shown. Journal of Investigative Dermatology  , DOI: ( /sj.jid ) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

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