Open AccessROS mediates interferon gamma induced phosphorylation of Src, through the Raf/ERK pathway, in MCF‐7 human breast cancer cell line
Author(s) -
Zibara Kazem,
Zeidan Asad,
Bjeije Hassan,
Kassem Nouhad,
Badran Bassam,
ElZein Nabil
Publication year - 2017
Publication title -
journal of cell communication and signaling
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 44
eISSN - 1873-961X
pISSN - 1873-9601
DOI - 10.1007/s12079-016-0362-6
Subject(s) - phosphorylation , protein kinase b , p38 mitogen activated protein kinases , signal transduction , cancer research , microbiology and biotechnology , mcf 7 , proto oncogene tyrosine protein kinase src , mapk/erk pathway , cell signaling , reactive oxygen species , kinase , biology , chemistry , cancer cell , cancer , human breast , genetics
Interferon gamma (IFN‐ɣ) is a pleiotropic cytokine which plays dual contrasting roles in cancer. Although IFN‐ɣ has been clinically used to treat various malignancies, it was recently shown to have protumorigenic activities. Reactive oxygen species (ROS) are overproduced in cancer cells, mainly due to NADPH oxidase activity, which results into several changes in signaling pathways. In this study, we examined IFN‐ɣ effect on the phosphorylation levels of key signaling proteins, through ROS production, in the human breast cancer cell line MCF‐7. After treatment by IFN‐ɣ, results showed a significant increase in the phosphorylation of STAT1, Src, raf, AKT, ERK1/2 and p38 signaling molecules, in a time specific manner. Src and Raf were found to be involved in early stages of IFN‐ɣ signaling since their phosphorylation increased very rapidly. Selective inhibition of Src‐family kinases resulted in an immediate significant decrease in the phosphorylation status of Raf and ERK1/2, but not p38 and AKT. On the other hand, IFN‐ɣ resulted in ROS generation, through H 2 O 2 production, whereas pre‐treatment with the ROS inhibitor NAC caused ROS inhibition and a significant decrease in the phosphorylation levels of AKT, ERK1/2, p38 and STAT1. Moreover, pretreatment with a selective NOX1 inhibitor resulted in a significant decrease of AKT phosphorylation. Finally, no direct relationship was found between ROS production and calcium mobilization. In summary, IFN‐ɣ signaling in MCF‐7 cell line is ROS‐dependent and follows the Src/Raf/ERK pathway whereas its signaling through the AKT pathway is highly dependent on NOX1.