title: Endogenous cGMP-dependent protein kinase reverses EGF-induced MAPK/ERK signal transduction through phosphorylation of VASP at Ser239.
abstract: In our previous study, we demonstrated that type II cGMP-dependent protein kinase (PKG II) was expressed at lower levels in different human cancer cell lines and that exogenous PKG II inhibited epidermal growth factor (EGF)-induced MAPK/ERK signaling. In order to investigate its functions further in this signaling pathway, it is necessary to elucidate whether endogenous PKG has the same effect or not. This study aimed to investigate the possible inhibitory effect of endogenous PKG activity on EGF-induced MAPK/ERK signal transduction in human lung cancer cells and its mechanism. Human small cell lung carcinoma cells (SCLCs) were treated with the PKG-selective cGMP analog 8-pCPT-cGMP to activate endogenous PKG, EGF and cGMP followed by EGF, respectively. The results showed that increased endogenous PKG activity inhibited the EGF-induced phosphorylation of the epidermal growth factor receptor (EGFR) and the binding between Sos1 and Grb2. In addition, EGF-triggered Ras activation was reversed by increased endogenous PKG activity. While the EGF-induced phosphorylation of MEK and ERK were inhibited by increased endogenous PKG activity, there was a significant increase of phosphorylated vasodilator-stimulated phosphoprotein (p-VASP) at Ser239. Furthermore, we investigated whether endogenous PKG exerted its effects on EGF-induced MAPK/ERK signaling through phosphorylation of VASP at Ser239. Downregulation of the levels of p-VASP Ser239 by point mutation blocked the effects of endogenous PKG on EGF-induced MAPK/ERK signal transduction. The data shown here suggest that endogenous PKG reverses the EGF-induced MAPK/ERK signaling pathway by phosphorylating VASP at Ser239.
authors.affiliation: School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013;
authors.forename: Yan
authors.initials: Y
authors.lastname: Tao
authors.forename: Yin-Jie
authors.initials: YJ
authors.lastname: Gu
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Cao
authors.forename: Xiu-Juan
authors.initials: XJ
authors.lastname: Bian
authors.forename: Ting
authors.initials: T
authors.lastname: Lan
authors.forename: Jian-Rong
authors.initials: JR
authors.lastname: Sang
authors.forename: Lu
authors.initials: L
authors.lastname: Jiang
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Hai
authors.initials: H
authors.lastname: Qian
authors.forename: Yong-Chang
authors.initials: YC
authors.lastname: Chen