Cyclic Guanosine Monophosphate (cGMP)-Dependent Protein Kinase II Blocks Epidermal Growth Factor (EGF)/Epidermal Growth Factor Receptor (EGFR)-Induced Biological Effects on Osteosarcoma Cells
Dapeng Li, Ye Hua, Lu Jiang, Yonghui Huang, Jiawei Yue, Yan Wu, Yongchang Chen
(Department of Orthopaedics, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, China (mainland))
Med Sci Monit 2018; 24:1997-2002
The present work was performed to detect the potential inhibitory effect of cyclic guanosine monophosphate (cGMP)-dependent protein kinase II (PKG II) on epidermal growth factor (EGF) receptor-induced biological activity and related signal cascades in osteosarcoma cells.
MATERIAL AND METHODS: We transfected the osteosarcoma MG-63 cell line with an adenoviral vector encoding PKG II cDNA (Ad-PKGII) and incubated the transfected cells with 250 μM 8-pCPT-cGMP to activate the PKG II. We stimulated the MG-63 cells with100 ng/ml EGF, and then detected their proliferation using a CCK-8 assay. Transwell assay was used to examine MG-63 cell migration; and Western blot analysis was used to detect expression of matrix metalloproteinase 9 (MMP-9) and activation of ERK and Akt.
RESULTS: Stimulating cells by 100 ng/ml EGF promoted MG-63 cell proliferation and migration, ERK and Akt phosphorylation, and MMP-9 expression. These effects of EGF were inhibited in MG-63 cells infected with Ad-PKGII and incubated with 8-pCPT-cGMP.
CONCLUSIONS: Our results demonstrate that Ad-PKGII infection significantly inhibited EGF-induced proliferation and migration, as well as the associated-signal cascades; which indicates that PKG II might be a potential anti-cancer factor.
Keywords: Cell Proliferation, Cyclic GMP-Dependent Protein Kinase Type II, Epidermal Growth Factor, Osteosarcoma, Receptor, Epidermal Growth Factor, Transcellular Cell Migration