04 April 2018 : Laboratory Research
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 Li1AEG*, Ye Hua1DE, Lu Jiang2CF, Yonghui Huang1BD, Jiawei Yue1CE, Yan Wu3AEG, Yongchang Chen3DFDOI: 10.12659/MSM.905892
Med Sci Monit 2018; 24: LBR1997-2002
Abstract
BACKGROUND: 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: Cyclic GMP-Dependent Protein Kinase Type II, Epidermal Growth Factor, Receptor, Epidermal Growth Factor, Transcellular Cell Migration
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