The online version of this article (doi:10.1186/1476-4598-11-22) contains supplementary material, which is available to authorized users.
Erika von Euw, Mohammad Atefi contributed equally to this work.
Erika von Euw, Mohammad Atefi, Narsis Attar, Connie Chu, Sybil Zachariah, Barry L. Burgess, Stephen Mok, Charles Ng, Deborah J.L. Wong, Bartosz Chmielowski, Richard C. Koya, Tara A. McCannel: Have no competing interests. David I. Lichter, Elena Izmailova: Are employees of Millennium Pharmaceuticals, Inc., the manufacturer of TAK733. Antoni Ribas: Is a compensated ad hoc advisor to Millennium Pharmaceuticals, Inc., the manufacturer of TAK733.
EE, MA, NA, CC, SZ, BLB, SM, CN, DJLW, DIL, RCK: Performed experiments. EE, MA, TAM, EI, AR: Designed the studies. BLB, BC, EI, AR: Provided key reagents. EE, MA, AR: Wrote the manuscript. EE, MA, NA, CC, SZ, BLB, SM, CN, DJLW, BC, DIL, RCK, TAM, EI, AR: All authors approved the final manuscript.
TAK733 is a novel allosteric, non-ATP-binding, inhibitor of the BRAF substrates MEK-1/2.
The growth inhibitory effects of TAK733 were assessed in a panel of 27 cutaneous and five uveal melanoma cell lines genotyped for driver oncogenic mutations. Flow cytometry, Western blots and metabolic tracer uptake assays were used to characterize the changes induced by exposure to TAK733.
Fourteen cutaneous melanoma cell lines with different driver mutations were sensitive to the antiproliferative effects of TAK733, with a higher proportion of BRAF V600E mutant cell lines being highly sensitive with IC50s below 1 nM. The five uveal melanoma cell lines had GNAQ or GNA11 mutations and were either moderately or highly sensitive to TAK733. The tested cell lines wild type for NRAS, BRAF, GNAQ and GNA11 driver mutations were moderately to highly resistant to TAK733. TAK733 led to a decrease in pERK and G1 arrest in most of these melanoma cell lines regardless of their origin, driver oncogenic mutations and in vitro sensitivity to TAK733. MEK inhibition resulted in increase in pMEK more prominently in NRAS Q61L mutant and GNAQ mutant cell lines than in BRAF V600E mutant cell lines. Uptake of the metabolic tracers FDG and FLT was inhibited by TAK733 in a manner that closely paralleled the in vitro sensitivity assays.
The MEK inhibitor TAK733 has antitumor properties in melanoma cell lines with different oncogenic mutations and these effects could be detectable by differential metabolic tracer uptake.
Additional file 1 : Figure S1TAK733 MTS-based colorimetric cell proliferation assay curves in melanoma cell lines of cutaneous origin according to their BRAF (A) or NRAS (B) mutational status, WT (C) and of uveal origin (D). Modulation of the melanoma cell line viability at a range of different concentrations of TAK733. The effects of TAK733 on cell growth and viability were analyzed after 72 hours of treatment using an MTS assay. (PPT 245 KB)12943_2011_1023_MOESM1_ESM.ppt
Additional file 2 : Figure S2 Time-course analyses of the effects of TAK733 on the signaling of the MAPK and PI3K/AKT pathways by Western blot. Two BRAF V600E melanoma cell lines were exposed for varying time points to TAK733. A) The sensitive BRAF V600E mutated cutaneous melanoma cell line M229; B) The resistant BRAF V600E mutated cutaneous melanoma cell line M233. (PPT 2 MB)12943_2011_1023_MOESM2_ESM.ppt
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- Antitumor effects of the investigational selective MEK inhibitor TAK733 against cutaneous and uveal melanoma cell lines
Erika von Euw
Barry L Burgess
Deborah JL Wong
David I Lichter
Richard C Koya
Tara A McCannel
- BioMed Central
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