Loss of Spry1 reduces growth of BRAFV600-mutant cutaneous melanoma and improves response to targeted therapy

摘要

, Box plots showing the expression of Spry1 gene in normal tissues, and in primary and metastatic CM considering data taken from UALCAN Database ( ), and in primary and metastatic CM for selected experiments taken from HCMDB Database ( ). Statistically significant differences were indicated: *  c OncoPrint showing Spry1 gene expression in BRAF -mutant CM samples taken from TCGA. mRNA expression of Spry1 was analyzed by qRT-PCR assay in six BRAF -mutant CM cell lines, and normalized to β-actin. Each bar represents  = 2 biological replicates, three technical replicates each; mean ± standard deviation (SD). The levels of Spry1, phospho-ERK1/2 (pERK1/2), and total ERK1/2 protein were determined by western blot in six BRAF -mutant CM cell lines. β-Tubulin was used as a loading control. Western blot images are representative of three independent experiments. Cytoplasmic localization of Spry1 in Mel 599 and Mel 611 CM cell lines. 3 × 10 cells were acquired and analyzed with the ImageStreamX instrument. , Mel 611 cells were treated or not (NT) with the MEKi U0126 (50 μmol/L) for 2, 4, and 8 h, and then the expression of SPRY1 was determined by qRT-PCR ( ) and western blot analyses ( ). Each bar represents  = 3 biological replicates, 3 technical replicates each; means ± SD. Statistically significant differences were indicated: **