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ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma.

Smit MA, Maddalo G, Greig K, Raaijmakers LM, Possik PA, van Breukelen B, Cappadona S, Heck AJ, Altelaar AF, Peeper DS - Mol. Syst. Biol. (2014)

Bottom Line: However, most patients eventually relapse due to drug resistance.We found many proteins to be induced upon PLX4720 (BRAF inhibitor) treatment that are known to be involved in BRAF inhibitor resistance, including FOXD3 and ErbB3.Several proteins were down-regulated, including Rnd3, a negative regulator of ROCK1 kinase.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis confirms deactivation of the MAPK pathway and MS-based phosphosite and protein quantificationCells were plated and treated the next day with either DMSO as a control (ctr) or 0.5 μM PLX4720 for 1 (1d) and 3 days (3d) and analyzed for proteins as indicated. β-actin serves as a loading control.MEK, ERK and p90RSK phosphorylation levels.RPS6 phosphorylation levels.Expression levels of several MS-quantified proteins.
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fig03: Western blot analysis confirms deactivation of the MAPK pathway and MS-based phosphosite and protein quantificationCells were plated and treated the next day with either DMSO as a control (ctr) or 0.5 μM PLX4720 for 1 (1d) and 3 days (3d) and analyzed for proteins as indicated. β-actin serves as a loading control.MEK, ERK and p90RSK phosphorylation levels.RPS6 phosphorylation levels.Expression levels of several MS-quantified proteins.

Mentions: As expected, exposure to PLX4720 led to down-regulation of the phosphorylation state of kinases within the MAPK pathway: phospho-MEK, phospho-ERK1/2 and phospho-p90RSK (Fig3A). Indeed also in our mass spectrometry analysis, we observed strong down-regulation of the phosphorylation states of ERK1 (at 1 and 3 days) and ERK2 (at 3 days; Supplementary Table S1), which indicate a responsive state of the employed cell line. Moreover, the phosphorylation state of RPS6 was significantly down-regulated especially at 1 day, indicative of an inactive state of the mTORC1 pathway (Fig3B, Supplementary Table S1) (Roux et al, 2007).


ROCK1 is a potential combinatorial drug target for BRAF mutant melanoma.

Smit MA, Maddalo G, Greig K, Raaijmakers LM, Possik PA, van Breukelen B, Cappadona S, Heck AJ, Altelaar AF, Peeper DS - Mol. Syst. Biol. (2014)

Western blot analysis confirms deactivation of the MAPK pathway and MS-based phosphosite and protein quantificationCells were plated and treated the next day with either DMSO as a control (ctr) or 0.5 μM PLX4720 for 1 (1d) and 3 days (3d) and analyzed for proteins as indicated. β-actin serves as a loading control.MEK, ERK and p90RSK phosphorylation levels.RPS6 phosphorylation levels.Expression levels of several MS-quantified proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300494&req=5

fig03: Western blot analysis confirms deactivation of the MAPK pathway and MS-based phosphosite and protein quantificationCells were plated and treated the next day with either DMSO as a control (ctr) or 0.5 μM PLX4720 for 1 (1d) and 3 days (3d) and analyzed for proteins as indicated. β-actin serves as a loading control.MEK, ERK and p90RSK phosphorylation levels.RPS6 phosphorylation levels.Expression levels of several MS-quantified proteins.
Mentions: As expected, exposure to PLX4720 led to down-regulation of the phosphorylation state of kinases within the MAPK pathway: phospho-MEK, phospho-ERK1/2 and phospho-p90RSK (Fig3A). Indeed also in our mass spectrometry analysis, we observed strong down-regulation of the phosphorylation states of ERK1 (at 1 and 3 days) and ERK2 (at 3 days; Supplementary Table S1), which indicate a responsive state of the employed cell line. Moreover, the phosphorylation state of RPS6 was significantly down-regulated especially at 1 day, indicative of an inactive state of the mTORC1 pathway (Fig3B, Supplementary Table S1) (Roux et al, 2007).

Bottom Line: However, most patients eventually relapse due to drug resistance.We found many proteins to be induced upon PLX4720 (BRAF inhibitor) treatment that are known to be involved in BRAF inhibitor resistance, including FOXD3 and ErbB3.Several proteins were down-regulated, including Rnd3, a negative regulator of ROCK1 kinase.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.

No MeSH data available.


Related in: MedlinePlus