Journal: Genome Medicine

Article Title: The expressed mutational landscape of microsatellite stable colorectal cancers

doi: 10.1186/s13073-021-00955-2

Figure Lengend Snippet: Correlation between TP53 and RAS / BRAF V600E mutation expression levels and sensitivity to targeted anticancer agents in pre-clinical models. a Sensitivity to the EGFR inhibitor erlotinib, the MEK inhibitor trametinib, and the MDM2 inhibitor idasanutlin in a panel of 29 unique CRC cell lines plotted according to RAS / BRAF V600E or TP53 mutation status, as indicated (mut, mutated; wt, wild-type; color codes are shown in c ). Higher DSS indicates stronger sensitivity. p -value is from Welch’s t -test of wild-type versus mutated samples. b , c Upper panels show the mutation status for RAS / BRAF V600E and TP53 in each of the 7 selected cell lines and 8 patient-derived organoids (PDOs). Scatter plots show the DSS of matched drugs versus mutant allele expression levels (color-coded as indicated). Spearman’s correlations in blue are for KRAS -mutated PDOs only (excluding the single NRAS -mutated sample). d Scatter plot of RNA-level versus DNA-level MAFs of RAS and TP53 in matched primary and metastatic tumor samples from each of four patients (three with RAS mutations and two with TP53 mutations). Patient 2 showed higher relative expression of the RAS mutant allele in the metastasis

Article Snippet: The MDM2-TP53 inhibitor idasanutlin (MedChemExpress, Monmouth Junction, NJ, USA), three EGFR inhibitors (afatinib: Selleck Chemicals; erlotinib: MedChemExpress; and lapatinib: LC Laboratories, Woburn, MA, USA) and two MEK inhibitors (binimetinib and trametinib, ChemieTek, Indianapolis, IN, USA) were included in the screens at five and nine different concentrations over a 10,000-fold concentration range each (typically 1–10,000 nmol/L) in the cell lines and PDOs, respectively.

Techniques: Mutagenesis, Expressing, Derivative Assay

Journal: Journal of Personalized Medicine

Article Title: GPR30 Activation by 17β-Estradiol Promotes p62 Phosphorylation and Increases Estrogen Receptor α Protein Expression by Inducing Its Release from a Complex Formed with KEAP1

doi: 10.3390/jpm11090906

Figure Lengend Snippet: GPR30 activation by 17β-estradiol elicits the SRC/EGFR/PI3K/Akt/mTOR signaling pathway and promotes p62 phosphorylation—which in turn induces ESR1 protein expression through estrogen receptor α (ESR1) detachment from a complex formed with KEAP1. This complex interplay provides a link between the rapid-signal effects of estrogens (mediated by GRP30) and their canonical genomic effects (mediated by ESR1).

Article Snippet: To explore the impact of the pharmacological modulation of different signaling pathways after exposure to 17β-estradiol, cells were pretreated with a proteasome inhibitor (MG132; Sigma-Aldrich, St. Louis, MO, USA; concentration: 5 μM), a protein synthesis inhibitor (cycloheximide; Sigma-Aldrich; concentration: 25 μM), a GRP30 agonist (G1; Tocris Bioscience, Bristol, UK; concentration: 100 nM), a GRP30 antagonist (G15; Tocris Bioscience, concentration: 100 nM), 17β-estradiol (Sigma-Aldrich; concentration: 100 nM), a PI3K inhibitor (LY294002; Sigma-Aldrich; concentration: 1 μM), an EGFR inhibitor (lapatinib; GlaxoSmithKline plc, London, UK, concentration: 120 nM), an SRC inhibitor (Sigma-Aldrich; concentration: 1 μM), an mTOR inhibitor that activates autophagy (everolimus; Novartis Pharma AG, Basel, Switzerland; concentration: 1 μM), a PKC inhibitor (sotrastaurin; MedChemExpress; concentration: 1 μM), a TAK inhibitor (takinib; MedChemExpress; concentration: 1 μM), a casein kinase inhibitor (PF-670462; MedChemExpress; concentration: 1 μM), and an autophagy inhibitor (chloroquine; Sigma-Aldrich; concentration: 10 μM).

Techniques: Activation Assay, Phospho-proteomics, Expressing