Mechanistic Analysis of an Extracellular Signal–Regulated Kinase 2–Interacting Compound that Inhibits Mutant BRAF-Expressing Melanoma Cells by Inducing Oxidative Stress
العنوان: | Mechanistic Analysis of an Extracellular Signal–Regulated Kinase 2–Interacting Compound that Inhibits Mutant BRAF-Expressing Melanoma Cells by Inducing Oxidative Stress |
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المؤلفون: | Maureen A. Kane, Ramin Samadani, Lijia Chen, Ramon Martinez, Ivie L. Conlon, Bryan Mackowiak, Paul Shapiro, Kellie Hom, Steven Fletcher, Garrick Centola, Weiliang Huang |
المصدر: | The Journal of Pharmacology and Experimental Therapeutics |
بيانات النشر: | The American Society for Pharmacology and Experimental Therapeutics, 2021. |
سنة النشر: | 2021 |
مصطلحات موضوعية: | 0301 basic medicine, MAPK/ERK pathway, Proto-Oncogene Proteins B-raf, MAP Kinase Signaling System, Antineoplastic Agents, Jurkat cells, Proto-Oncogene Mas, 03 medical and health sciences, Drug Discovery and Translational Medicine, Jurkat Cells, 0302 clinical medicine, Catalytic Domain, Humans, Binding site, Transcription factor, Melanoma, Cell Proliferation, Pharmacology, Mitogen-Activated Protein Kinase 1, Chemistry, Activator (genetics), Kinase, Cell growth, Cell biology, Oxidative Stress, 030104 developmental biology, Cancer cell, Molecular Medicine, 030217 neurology & neurosurgery, HeLa Cells, Protein Binding |
الوصف: | Constitutively active extracellular signal-regulated kinase (ERK) 1/2 signaling promotes cancer cell proliferation and survival. We previously described a class of compounds containing a 1,1-dioxido-2,5-dihydrothiophen-3-yl 4-benzenesulfonate scaffold that targeted ERK2 substrate docking sites and selectively inhibited ERK1/2-dependent functions, including activator protein-1-mediated transcription and growth of cancer cells containing active ERK1/2 due to mutations in Ras G-proteins or BRAF, Proto-oncogene B-RAF (Rapidly Acclerated Fibrosarcoma) kinase. The current study identified chemical features required for biologic activity and global effects on gene and protein levels in A375 melanoma cells containing mutant BRAF (V600E). Saturation transfer difference-NMR and mass spectrometry analyses revealed interactions between a lead compound (SF-3-030) and ERK2, including the formation of a covalent adduct on cysteine 252 that is located near the docking site for ERK/FXF (DEF) motif for substrate recruitment. Cells treated with SF-3-030 showed rapid changes in immediate early gene levels, including DEF motif-containing ERK1/2 substrates in the Fos family. Analysis of transcriptome and proteome changes showed that the SF-3-030 effects overlapped with ATP-competitive or catalytic site inhibitors of MAPK/ERK Kinase 1/2 (MEK1/2) or ERK1/2. Like other ERK1/2 pathway inhibitors, SF-3-030 induced reactive oxygen species (ROS) and genes associated with oxidative stress, including nuclear factor erythroid 2-related factor 2 (NRF2). Whereas the addition of the ROS inhibitor N-acetyl cysteine reversed SF-3-030-induced ROS and inhibition of A375 cell proliferation, the addition of NRF2 inhibitors has little effect on cell proliferation. These studies provide mechanistic information on a novel chemical scaffold that selectively regulates ERK1/2-targeted transcription factors and inhibits the proliferation of A375 melanoma cells through a ROS-dependent mechanism. SIGNIFICANCE STATEMENT: Constitutive activation of the extracellular signal-regulated kinase (ERK1/2) pathway drives the proliferation and survival of many cancer cell types. Given the diversity of cellular functions regulated by ERK1/2, the current studies have examined the mechanism of a novel chemical scaffold that targets ERK2 near a substrate binding site and inhibits select ERK functions. Using transcriptomic and proteomic analyses, we provide a mechanistic basis for how this class of compounds inhibits melanoma cells containing mutated BRAF and active ERK1/2. |
اللغة: | English |
تدمد: | 1521-0103 0022-3565 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::54b230868d4274902242e0aeb5922c33 http://europepmc.org/articles/PMC7788356 |
حقوق: | OPEN |
رقم الأكسشن: | edsair.doi.dedup.....54b230868d4274902242e0aeb5922c33 |
قاعدة البيانات: | OpenAIRE |
تدمد: | 15210103 00223565 |
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