Src Kinase Is Biphosphorylated at Y416/Y527 and Activates the CUB-Domain Containing Protein 1/Protein Kinase C ? Pathway in a Subset of Triple-Negative Breast Cancers.

ABSTRACT: Targeted therapeutics are needed for triple-negative breast cancer (TNBC). In this study, we investigated the activation of Src family of cytoplasmic tyrosine kinases (SFKs) and two SFK substrates-CUB-domain containing protein 1 (CDCP1) and protein kinase C ? (PKC?)-in 56 formalin-fixed, paraffin-embedded (FFPE) TNBCs. Expression of SFK phosphorylated at Y416 (SFK_pY416⁺) in tumor cells was strongly associated with phosphorylation of CDCP1 and PKC? (CDCP1_ pY743⁺ and PKC?_pY311⁺), as assessed by immunohistochemistry, indicating increased SFK activity in situ. To enable biochemical analysis, protein extraction from FFPE tissue was optimized. Cleaved CDCP1 isoform (70 kDa) was expressed to a varying degree in all samples but only phosphorylated in TNBC tumor cells that expressed SFK_pY416. Interestingly, active SFK was found to be biphosphorylated (SFK_pY416⁺/pY527⁺). Biphosphorylated active SFK was observed more frequently in forkhead box protein A1 (FOXA1)^- TNBCs. In addition, in SFK_pY416^- samples, FOXA1⁺ TNBC tended to be SFK_pY527⁺ (classic inactive SFK), and FOXA1^- TNBC tended to be SFK_pY527^- (SFK poised for activation). Strong SFK_pY416 staining was also observed in tumor-infiltrating lymphocytes in a subset of TNBCs with high tumor-infiltrating lymphocyte content. This report will facilitate protein biochemical analysis of FFPE tumor samples and justifies the development of therapies targeting the SFK/CDCP1/PKC? pathway for TNBC treatment.

SUBMITTER: Nelson LJ 

PROVIDER: S-EPMC6983918 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature