Dual mTOR/DNA-PK Inhibitor CC-115 Induces Cell Death in Melanoma Cells and Has Radiosensitizing Potential

CC-115 is a dual inhibitor of the mechanistic target of rapamycin (mTOR) kinase and the DNA-dependent protein kinase (DNA-PK) that is currently being studied in phase I/II clinical trials. DNA-PK is essential for the repair of DNA-double strand breaks (DSB). Radiotherapy is frequently used in the palliative treatment of metastatic melanoma patients and induces DSBs. Melanoma cell lines and healthy-donor skin fibroblast cell lines were treated with CC-115 and ionizing irradiation (IR). Apoptosis, necrosis, and cell cycle distribution were analyzed. Colony forming assays were conducted to study radiosensitizing effects. Immunofluorescence microscopy was performed to determine the activity of homologous recombination (HR). In most of the malign cell lines, an increasing concentration of CC-115 resulted in increased cell death. Furthermore, strong cytotoxic effects were only observed in malignant cell lines. Regarding clonogenicity, all cell lines displayed decreased survival fractions during combined inhibitor and IR treatment and supra-additive effects of the combination were observable in 5 out of 9 melanoma cell lines. CC-115 showed radiosensitizing potential in 7 out of 9 melanoma cell lines, but not in healthy skin fibroblasts. Based on our data CC-115 treatment could be a promising approach for patients with metastatic melanoma, particularly in the combination with radiotherapy.

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Suppression subtractive hybridization profiles of radial growth phase and metastatic melanoma cell lines reveal novel potential targets

BMC Cancer ◽

10.1186/1471-2407-8-19 ◽

2008 ◽

Vol 8 (1) ◽

Cited By ~ 14

Author(s):

Josane F Sousa ◽

Enilza M Espreafico

Keyword(s):

Metastatic Melanoma ◽

Cell Lines ◽

Suppression Subtractive Hybridization ◽

Melanoma Cell ◽

Growth Phase ◽

Radial Growth ◽

Subtractive Hybridization ◽

Metastatic Melanoma Cell ◽

Melanoma Cell Lines

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Photodynamic treatment induces cell death by apoptosis or autophagy depending on the melanin content in two B16 melanoma cell lines

Oncology Reports ◽

10.3892/or.2012.2190 ◽

2012 ◽

Vol 29 (3) ◽

pp. 1196-1200 ◽

Cited By ~ 24

Author(s):

AGNÈS SPARSA ◽

SOLENN BELLATON ◽

THOMAS NAVES ◽

MARIE-ODILE JAUBERTEAU ◽

JEAN-MARIE BONNETBLANC ◽

...

Keyword(s):

Cell Death ◽

Cell Lines ◽

Melanoma Cell ◽

B16 Melanoma ◽

Melanin Content ◽

Photodynamic Treatment ◽

B16 Melanoma Cell ◽

Melanoma Cell Lines

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PAX3 knockdown in metastatic melanoma cell lines does not reduce MITF expression

Melanoma Research ◽

10.1097/cmr.0b013e328341c7e0 ◽

2011 ◽

Vol 21 (1) ◽

pp. 24-34 ◽

Cited By ~ 11

Author(s):

Shujie He ◽

Caiyun G. Li ◽

Lynn Slobbe ◽

Amy Glover ◽

Elaine Marshall ◽

...

Keyword(s):

Metastatic Melanoma ◽

Cell Lines ◽

Melanoma Cell ◽

Metastatic Melanoma Cell ◽

Mitf Expression ◽

Melanoma Cell Lines

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Inhibition of glucose metabolism through treatment of BRAF mutated metastatic melanoma with vemurafenib.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e21005 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e21005-e21005 ◽

Cited By ~ 1

Author(s):

Govind Warrier ◽

Lilibeth Lanceta ◽

Yoannis Imbert-Fernandez ◽

Jason Alan Chesney

Keyword(s):

Glucose Metabolism ◽

Protein Expression ◽

Metastatic Melanoma ◽

Cell Lines ◽

Melanoma Cell ◽

Control Point ◽

Braf Inhibitor ◽

Growth And Survival ◽

Metastatic Melanoma Cell ◽

Melanoma Cell Lines

e21005 Background: Increased glucose metabolism is a hallmark of neoplastic cells that allows self-promotion of growth and survival. The enzyme 6-phosphofructo-2-kinase (PFKFB3) is an integral controller of glycolysis by promoting the synthesis of fructose 2,6-bisphosphonate (F2,6BP) which activates 6-phoshofructo-1-kinase (PFK-1), a rate-limiting enzyme and essential control point in the glycolytic pathway. Additionally, mitogen-activated protein kinase (MAPK) is a key signaling pathway in a number of cancers with mutations of the BRAF component, described most commonly in melanoma, resulting in constitutive activation of the MAPK pathway. We aim to demonstrate that vemurafenib, a BRAF inhibitor, has antiglycolytic activity in sensitive melanoma cell lines which may help guide development of future therapies with specific attention to PFKFB3 as a potential enzymatic target to decrease glycolytic flux thereby inhibiting tumor growth and survival. Methods: Vemurafenib sensitive and resistant variants of two separate human metastatic melanoma cell lines (451Lu and WM983) were treated with 3 mM Vemurafenib for 24 and 48 hours. Additionally, cells from aforementioned lines were probed for PFKFB3 after 24 hours of treatment with vemurafenib. Glycolysis was measured by incubating cells in complete media containing 1 mCi [5-3H]glucose for 60 minutes. [3H]H2O produced by glycolysis through enolase was measured. Results: A decrease in PFKFB3 protein expression was found in vemurafenib sensitive cells compared to controls but not in resistant cells after 24h treatment with 3 mM vemurafenib in both 451Lu and WM983 metastatic melanoma cell lines (n = 2). Treatment with vemurafenib led to decrease in glycolysis compared to untreated controls in both vemurafenib sensitive metastatic melanoma cell lines but not in resistant cell lines (n = 5). Additionally, there was a significant reduction in glycolysis in vemurafenib resistant WM983 at 48 hours compared to resistant untreated control. Conclusions: BRAF mutated metastatic melanoma cells showed decrease in PFKFB3 protein expression and decreased glycolysis after treatment with BRAF inhibitor vemurafenib. Future studies will focus on assessing metastatic melanoma cell viability and glycolytic activity after treatment with combination BRAF inhibition and PFKFB3 specific inhibition.

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