Phenotypic characterization of the novel, non‑hotspot oncogenic KRAS mutants E31D and E63K

摘要: KRAS proto-oncogene, GTPase (KRAS) functions as a molecular switch at the apex of multiple signaling pathways controlling cell proliferation, differentiation, migration, and survival. Canonical mutants, such those in codons 12 13, produce constitutively active oncoproteins that short-circuit epidermal growth factor receptor (EGFR)-initiated signaling, resulting dysregulated downstream effectors associated with cellular transformation. Therefore, anti-EGFR therapy provides little to no clinical benefit patients activating mutations. Current genotyping procedures based on canonical mutation detection only account for ~40% non-responders, highlighting need identify additional predictive biomarkers. In present study, two novel non-hotspot mutations were functionally characterized vitro: E31D was identified from genetic screen colorectal cancer specimens UP-National Institutes Health. E63K is curated Catalogue Somatic Mutations Cancer database. Similar mutants G12D G13D, NIH3T3 cells overexpressing showed altered morphology characteristically smaller, rounder, highly refractile compared their non-transformed counterparts. Filamentous actin staining also indicated cytoplasmic shrinkage, membrane ruffling, formation pseudopod protrusions. Further, they displayed higher proliferative rates migratory scratch wound assays negative controls. These empirical findings suggest impact mutations, which may contribute resistance therapy. Complementary studies elucidate mechanisms underlying transforming effect rare are required. parallel, oncogenic capacity vivo should be investigated.