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High‐risk endometrial cancer proteomic profiling reveals that FBXW7 mutation alters L1CAM and TGM2 protein levels
Author(s) -
Urick Mary Ellen,
Yu EunJeong,
Bell Daphne W.
Publication year - 2021
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/cncr.33567
Subject(s) - crispr , biology , cancer research , microbiology and biotechnology , l1 , genotype , cell culture , endometrial cancer , cell , mutation , genetics , cancer , gene
Background FBXW7 is frequently somatically mutated in grade 3 endometrioid endometrial cancers (G3EECs) and serous endometrial cancers (SECs), which are high‐risk cancers associated with poor outcomes and in need of novel treatment options. The aim of this study was to determine the proteomic effects of 3 FBXW7 mutations in high‐risk endometrial cancers (ECs). Methods Clustered regularly interspaced short palindromic repeats (CRISPR) editing was used to generate 3 HEC‐50B G3EEC derivative cell lines, each of which harbored 1 FBXW7 mutation, and to revert an endogenous FBXW7 mutation in HEC‐1‐B grade 2 endometrioid endometrial cancer (G2EEC) cells to the wild‐type genotype. Proteomic profiling based on liquid chromatography–tandem mass spectrometry was used to determine protein differences between the HEC‐50B derivative lines and parental cells. Western blot analysis was performed to assess differential protein levels of CRISPR‐edited derivative lines originating from HEC‐50B, ARK1 (SEC), ARK4 (SEC), HEC‐1‐B, and JHUEM‐1 (G2EEC) cell lines in comparison with parental cells. Results Results of this study demonstrated the effects of FBXW7 mutations on the proteome and phosphoproteome of HEC‐50B G3EEC cells and highlighted proteins that also exhibited altered levels in FBXW7 ‐mutated ARK1 and ARK4 SEC cells, including 2 potentially druggable proteins: L1 cell adhesion molecule (L1CAM) and transglutaminase 2 (TGM2). Furthermore, they demonstrated that reversion of an endogenous FBXW7 mutation to the wild‐type genotype in JHUEM‐1 and HEC‐1‐B G2EEC cells resulted in decreased L1CAM and TGM2 protein levels. Conclusions L1CAM and TGM2 protein levels are affected by FBXW7 mutations in ECs.