Preclinical evaluation of dasatinib, a potent Src kinase inhibitor, in melanoma cell lines - PubMed (original) (raw)

Preclinical evaluation of dasatinib, a potent Src kinase inhibitor, in melanoma cell lines

Alex J Eustace et al. J Transl Med. 2008.

Abstract

Background: Metastatic melanoma is a highly chemotherapy resistant tumour. The use of newer targeted therapies alone and in combination with chemotherapy may offer new hope of improving response to treatment. Dasatinib, a multi-target kinase inhibitor, is currently approved for the treatment of chronic myeloid leukaemia and has shown promising results in preclinical studies in a number of solid tumours.

Methods: We examined the effects of dasatinib on proliferation, chemo-sensitivity, cell cycle arrest, apoptosis, migration and invasion in human melanoma cell lines. Expression and activation of Src kinase, FAK and EphA2 were also examined in the melanoma cells.

Results: Dasatinib inhibited growth of three of the five melanoma cell lines. Comparison with sorafenib showed that in these three cell lines dasatinib inhibited growth at lower concentrations than sorafenib. Dasatinib in combination with the chemotherapy drug temozolomide showed greater efficacy than either drug alone. Dasatinib induced cell cycle arrest and apoptosis and significantly inhibited cell migration and invasion of melanoma cells. Dasatinib inhibition of proliferation was associated with reduced phosphorylation of Src kinase, while decreased phosphorylation of FAK was implicated in dasatinib-mediated inhibition of migration and invasion in melanoma cells.

Conclusion: Dasatinib has both anti-proliferative and anti-invasive effects in melanoma cells and combined with chemotherapy may have clinical benefit in the treatment of malignant melanoma.

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Figures

Figure 1

Figure 1

Percentage growth inhibition by A) dasatinib and B) sorafenib in a panel of melanoma cell lines. Error bars represent the standard deviation of triplicate experiments.

Figure 2

Figure 2

Combination assays testing dasatinib with temozolomide at the specified ratios in (A) HT144 (ratio 1:1500), (B) Lox-IMVI (ratio 1:3000), (C) Malme-3M (ratio 1:800) and (D) Sk-Mel-28 (ratio 1:800) cells. Concentrations of temozolomide are represented as a ratio of the dasatinib concentration. Error bars represent the standard deviation of triplicate experiments.

Figure 3

Figure 3

Measurement of dasatinib induced apoptosis in HT144, Lox-IMVI and Malme-3M using the TUNEL assay.

Figure 4

Figure 4

Effect of dasatinib on (A) invasion and (B) migration in HT144 and Sk-Mel-28 melanoma cell lines. Error bars represent the standard deviation of triplicate assays. '*' indicates p < 0.05.

Figure 5

Figure 5

Western blotting for Src kinase, phospho-Src kinase py 418, FAK, phospho-FAK py 397 and py 861, immunoprecipitated (IP) EphA2, IP phospho-EphA2 and α-tubulin in (A) the panel of melanoma cell lines; and (B) HT144, Lox-IMVI, Malme-3M, Sk-Mel-5 and Sk-Mel-28 untreated (control) or treated with 100 nM dasatinib for 6 hours. (C) Western blotting for IP EphA2, IP phospho-EphA2 in Lox-IMVI untreated (control) and treated with 100 nM dasatinib for up to 48 hours.

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References

    1. Gogas HJ, Kirkwood JM, Sondak VK. Chemotherapy for metastatic melanoma: time for a change? Cancer. 2007;109:455–464. doi: 10.1002/cncr.22427. - DOI - PubMed
    1. Quirt I, Verma S, Petrella T, Bak K, Charette M. Temozolomide for the treatment of metastatic melanoma: a systematic review. Oncologist. 2007;12:1114–1123. doi: 10.1634/theoncologist.12-9-1114. - DOI - PubMed
    1. Sharma A, Trivedi NR, Zimmerman MA, Tuveson DA, Smith CD, Robertson GP. Mutant V599EB-Raf regulates growth and vascular development of malignant melanoma tumors. Cancer Res. 2005;65:2412–2421. doi: 10.1158/0008-5472.CAN-04-2423. - DOI - PubMed
    1. Eisen T, Ahmad T, Flaherty KT, Gore M, Kaye S, Marais R, Gibbens I, Hackett S, James M, Schuchter LM, et al. Sorafenib in advanced melanoma: a Phase II randomised discontinuation trial analysis. Br J Cancer. 2006;95:581–586. doi: 10.1038/sj.bjc.6603291. - DOI - PMC - PubMed
    1. Flaherty KT. Sorafenib: delivering a targeted drug to the right targets. Expert Rev Anticancer Ther. 2007;7:617–626. doi: 10.1586/14737140.7.5.617. - DOI - PubMed

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