The phospholipase D inhibitor FIPI potently blocks EGF-induced calcium signaling in human breast cancer cells
Autor: | Nadine Rommerswinkel, Silvia Keil, Bernd Niggemann, Thomas Dittmar, Sandina A. Gnoth, Helena M. Stricker |
---|---|
Rok vydání: | 2021 |
Předmět: |
0301 basic medicine
Indoles PLC-γ1 lcsh:Medicine chemistry.chemical_element Breast Neoplasms Calcium Biochemistry 03 medical and health sciences chemistry.chemical_compound Breast cancer 0302 clinical medicine Cell Movement Epidermal growth factor Cell Line Tumor Phospholipase D Humans Receptors sigma Cell migration Calcium Signaling Enzyme Inhibitors Phosphorylation RNA Small Interfering lcsh:QH573-671 skin and connective tissue diseases Molecular Biology Protein kinase B Calcium signaling Epidermal Growth Factor lcsh:Cytology Chemistry Research lcsh:R Cell Biology Phosphatidic acid Domperidone Cell biology ErbB Receptors 030104 developmental biology 030220 oncology & carcinogenesis Cancer cell Second messenger system Female PLD |
Zdroj: | Cell Communication and Signaling, Vol 19, Iss 1, Pp 1-16 (2021) Cell Communication and Signaling : CCS |
ISSN: | 1478-811X |
Popis: | BackgroundPhosphotyrosine kinase (PTK)-mediated phospholipase C-γ1 (PLC-γ1) signaling plays a crucial role in the release of the universal second messenger calcium from intracellular stores, which is mandatory for several cellular processes, including cell migration. However, PLC-γ1 could also be activated in a PTK-independent manner by phospholipase D (PLD)-derived phosphatidic acid (PA). Because both higher PLD expression levels and PLD activity have also been associated with breast cancer cell invasion and migration, we wondered whether there might be a link between PLD and PLC-γ1, which was investigated in this study.MaterialsMDA-MB-468-NEO (EGFR positive) and MDA-MB-468-HER2 (EGFR and HER2 positive) human breast cancer cells were used in this study. The migratory behavior of the cells in the presence of epidermal growth factor (EGF) and the PLD inhibitor 5-fluoro-2-indolyl-des-chlorohalopemide (FIPI) was analyzed using the 3D collagen matrix migration assay. Changes in cytosolic calcium levels in the presence of EGF, FIPI and Sig-1R agonists and antagonists as well as in PLD1 siRNA knockdown cells were determined by flow cytometry. Western blot analyses were performed to determine the basal expression levels and phosphorylation patterns of EGFR, HER2, AKT, MAPKp42/44, PLC-γ1 and Sig-1R.ResultsThe EGF-induced migration of MDA-MB-468-NEO and MDA-MB-468-HER2 cells was significantly impaired by FIPI. Likewise, FIPI also significantly abolished EGF-induced calcium release in both cell lines. However, neither the expression levels nor the phosphorylation patterns of EGFR, HER2, AKT, MAPKp42/44and PLC-γ1 were markedly changed by FIPI. Knockdown of PLD1 expression by siRNA also significantly impaired EGF-induced calcium release in both cell lines. Targeting Sig-1R, which interacts with IP3R, with the antagonist BD1047 also abrogated EGF-induced calcium release. However, EGF-induced calcium release was also impaired if cells were treated with the Sig-1R agonists PRE084 and PPBP maleate.ConclusionIn summary, blocking PLD activity with the specific inhibitor FIPI or knocking down PDL1 expression by siRNA significantly impaired EGF-induced calcium release in MDA-MB-468-NEO and MDA-MB-468-HER2 cells, likely indicating a connection between PLD activity and PLC-γ1-mediated calcium signaling. However, how PLD activity interferes with the release of calcium from intracellular stores remains unclear. |
Databáze: | OpenAIRE |
Externí odkaz: |