Suppression of mTOR complex 2-dependent AKT phosphorylation in melanoma cells by combined treatment with rapamycin and LY294002

• Published in: British journal of dermatology (1951) Vol. 160; no. 5; pp. 955 - 964
• Main Authors: Werzowa, J., Cejka, D., Fuereder, T., Dekrout, B., Thallinger, C., Pehamberger, H., Wacheck, V., Pratscher, B.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.05.2009; Wiley-Blackwell
• Subjects: Apoptosis - drug effects; Biological and medical sciences; Blotting, Western; Cell Cycle - drug effects; Cell Line, Tumor; Chromones - therapeutic use; Dermatology; Drug Therapy, Combination; Humans; LY294002; Medical sciences; melanoma; Melanoma - drug therapy; Melanoma - metabolism; Morpholines - therapeutic use; mTORC1; mTORC2; Phosphorylation - drug effects; Proto-Oncogene Proteins c-akt - drug effects; rapamycin; Reverse Transcriptase Polymerase Chain Reaction; Sirolimus - therapeutic use; Skin Neoplasms - drug therapy; Skin Neoplasms - metabolism; Transcription Factors - antagonists & inhibitors; Tumors of the skin and soft tissue. Premalignant lesions; Antineoplastic agent; Phosphorylation; LY294002; Sirolimus; Dermatology; melanoma; Malignant tumor; Antibiotic; Malignant melanoma; Combined treatment; Immunosuppressive agent; Antibacterial agent; mTORC2; mTORC1; Cancer; rapamycin
• ISSN: 0007-0963; 1365-2133
• DOI: 10.1111/j.1365-2133.2008.08991.x

Summary Background  Inhibition of mTOR complex 1 (mTORC1) with rapamycin leads to phosphorylation of AKT in some cancer cells, with unknown biological consequences. The role of this phosphorylation in melanoma is unknown, although preliminary clinical data indicate poor activity of rapalogues in melanoma. Objectives  We aimed at elucidating the role of AKT phosphorylation after mTORC1 inhibition in melanoma cells. Methods  Western blotting, apoptosis assays, cell cycle analyses and viability assays were performed to analyse the effects of rapamycin and LY294002 treatment on melanoma cells. For suppression of mTOR complex 2 (mTORC2) an siRNA directed against rictor was used. Results  Rapamycin showed limited effects on cell viability but resulted in strong and lasting AKT phosphorylation in melanoma cells. Combined PI3K/mTOR inhibition with LY294002 had pronounced effects on viability but also led to increased AKT phosphorylation after prolonged treatment. In contrast, combination of rapamycin plus LY294002 suppressed AKT phosphorylation. Suppression of AKT phosphorylation did not correlate with decreases in cell viability. Inhibition of mTORC2 led to reduced levels of phosphorylated AKT. Conclusions  mTORC1 inhibition with rapamycin and with LY294002 can lead to AKT phosphorylation in melanoma cells via mTORC2. Combination of rapamycin and LY294002 suppresses AKT phosphorylation but without significant effect on treatment efficacy.