Nitric oxide/cyclic guanosine monophosphate inducers sodium nitroprusside and L-arginine inhibit the proliferation of gastric cancer cells via the activation of type II cyclic guanosine monophosphate-dependent protein kinase

Yao,Xiaoyuan; Wu,Yan; Zhu,Miaolin; Qian,Hai; Chen,Yongchang

doi:10.3892/ol.2015.3229

Nitric oxide/cyclic guanosine monophosphate inducers sodium nitroprusside and L-arginine inhibit the proliferation of gastric cancer cells via the activation of type II cyclic guanosine monophosphate-dependent protein kinase

Authors:
- Xiaoyuan Yao
- Yan Wu
- Miaolin Zhu
- Hai Qian
- Yongchang Chen
View Affiliations

Affiliations: Department of Physiology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China
Published online on: May 19, 2015 https://doi.org/10.3892/ol.2015.3229
Pages: 479-484

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Nitric oxide (NO) may activate soluble guanylyl cyclase (sGC), resulting in the increase of intracellular cyclic guanosine monophosphate (cGMP), a key molecule in the activation of type II cGMP‑dependent protein kinase (PKG II). In our previous study, the membrane‑permeable cGMP analogue 8‑pCPT‑cGMP was used to activate PKG II. The aim of the present study was to investigate whether NO/sGC‑induced endogenous cGMP is able to activate PKG II and induce the corresponding effects. In the AGS gastric cancer cell line, the expression of PKG II was increased by infecting the cells with an adenoviral construct encoding PKG II cDNA (Ad‑PKG II) and the activation of PKG II was induced by 8‑pCPT‑cGMP (positive control), the NO donor sodium nitroprusside (SNP) and the NO precursor L‑arginine. ELISA was performed to detect the concentration of cGMP in AGS cells and the Cell Counting Kit‑8 was used to analyze the proliferation of differently treated cells. Western blot analysis was used to detect the expression and phosphorylation of associated proteins. The results demonstrated that the level of cGMP was increased in cells treated with the NO donor or precursor. There was an obvious increase of ser239 phosphorylation of the vasodilator‑stimulated phosphoprotein, representing the increase in the activity of PKG II. The epidermal growth factor (EGF)‑induced proliferation of AGS cells was inhibited by infection with Ad‑PKG II and treatment with SNP or L‑arginine. In addition, EGF‑induced tyrosine phosphorylation of the EGF receptor (EGFR) and tyrosine/serine phosphorylation of extracellular signal‑regulated kinase (ERK) were also inhibited by infection with Ad‑PKG II and treatment with the NO donor or precursor. These data indicated that NO donors and precursors may activate the expression of PKG II, thereby blocking EGF‑initiated signaling of the mitogen‑activated protein kinase/ERK pathway and inhibiting EGF‑induced proliferative activity through preventing the phosphorylation of EGFR at Tyr1068.

View Figures

View References

1	Bohme E and Schmidt HH: Nitric oxide and cytosolic guanylate cyclase: components of an intercellular signalling system. Z Kardiol 78 Suppl. 6:75–79. 1989.
2	Orstavik S, Natarajan V, Tasken K, Jahnsen T and Sandberg M: Characterization of the human gene encoding the type I alpha and type I beta cGMP-dependent protein kinase (PRKG1). Genomics. 42:311–318. 1997. View Article : Google Scholar : PubMed/NCBI
3	Orstavik S, Solberg R, Taskén K, et al: Molecular cloning, cDNA structure and chromosomal localization of the human type II cGMP-dependent protein kinase. Biochem Biophys Res Commun. 220:759–765. 1996. View Article : Google Scholar : PubMed/NCBI
4	Wells A: EGF receptor. Int J Biochem Cell Biol. 31:637–643. 1999. View Article : Google Scholar : PubMed/NCBI
5	Jiang H, Grenley MO, Bravo MJ, Blumhagen RZ and Edgar BA: EGFR/Ras/MAPK signaling mediates adult midgut epithelial homeostasis and regeneration in Drosophila. Cell Stem Cell. 8:84–95. 2011. View Article : Google Scholar : PubMed/NCBI
6	Chen YC, Ren F, Sang JR, Tao Y and Xu WR: Type II cGMP-dependent protein kinase inhibits proliferation of the gastric cancer cell line BGC-823. Mol Med Rep. 3:361–366. 2010. View Article : Google Scholar : PubMed/NCBI
7	Wu Y, Chen Y, Qu R, Lan T and Sang J: Type II cGMP-dependent protein kinase inhibits EGF-triggered signal transduction of the MAPK/ERK-mediated pathway in gastric cancer cells. Oncol Rep. 27:553–558. 2012.PubMed/NCBI
8	Lan T, Chen Y, Sang J, et al: Type II cGMP-dependent protein kinase inhibits EGF-induced MAPK/JNK signal transduction in breast cancer cells. Oncol Rep. 27:2039–2044. 2012.PubMed/NCBI
9	Jiang L, Lan T, Chen Y, et al: PKG II inhibits EGF/EGFR-induced migration of gastric cancer cells. PLoS One. 8:e616742013. View Article : Google Scholar : PubMed/NCBI
10	Medeiros JV, Gadelha GG, Lima SJ, et al: Role of the NO/cGMP/K (ATP) pathway in the protective effects of sildenafil against ethanol-induced gastric damage in rats. Br J Pharmacol. 153:721–727. 2008. View Article : Google Scholar : PubMed/NCBI
11	Russo I, Doronzo G, Mattiello L, De Salve A, Trovati M and Anfossi G: The activity of constitutive nitric oxide synthase is increased by the pathway cAMP/cAMP-activated protein kinase in human platelets. New insights into the antiaggregating effects of cAMP-elevating agents. Thromb Res. 114:265–273. 2004. View Article : Google Scholar : PubMed/NCBI
12	Kwiatkowski AV, Gertler FB and Loureiro JJ: Function and regulation of Ena/VASP proteins. Trends Cell Biol. 13:386–392. 2003. View Article : Google Scholar : PubMed/NCBI
13	Tao Y, Gu YJ, Cao ZH, et al: Endogenous cGMP-dependent protein kinase reverses EGF-induced MAPK/ERK signal transduction through phosphorylation of VASP at Ser239. Oncol Lett. 4:1104–1108. 2012.PubMed/NCBI
14	Wheeler S, Siwak DR, Chai R, et al: Tumor epidermal growth factor receptor and EGFR PY1068 are independent prognostic indicators for head and neck squamous cell carcinoma. Clin Cancer Res. 18:2278–2289. 2012. View Article : Google Scholar : PubMed/NCBI
15	Yamazaki T, Zaal K, Hailey D, Presley J, Lippincott-Schwartz J and Samelson LE: Role of Grb2 in EGF-stimulated EGFR internalization. J Cell Sci. 115:1791–1802. 2002.PubMed/NCBI
16	Rojas M, Yao S and Lin YZ: Controlling epidermal growth factor (EGF)-stimulated Ras activation in intact cells by a cell-permeable peptide mimicking phosphorylated EGF receptor. J Biol Chem. 271:27456–27461. 1996. View Article : Google Scholar : PubMed/NCBI
17	Saito T, Okada S, Ohshima K, et al: Differential activation of epidermal growth factor (EGF) receptor downstream signaling pathways by betacellulin and EGF. Endocrinology. 145:4232–4243. 2004. View Article : Google Scholar : PubMed/NCBI
18	Kolch W: Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J. 351:289–305. 2000. View Article : Google Scholar : PubMed/NCBI
19	Dumitrescu C, Biondi R, Xia Y, et al: Myocardial ischemia results in tetrahydrobiopterin (BH4) oxidation with impaired endothelial function ameliorated by BH4. Proc Natl Acad Sci USA. 104:15081–15086. 2007. View Article : Google Scholar : PubMed/NCBI
20	Knowles RG and Moncada S: Nitric oxide synthases in mammals. Biochem J. 298:249–258. 1994.PubMed/NCBI
21	Koesling D, Russwurm M, Mergia E, Mullershausen F and Friebe A: Nitric oxide-sensitive guanylyl cyclase: structure and regulation. Neurochem Int. 45:813–819. 2004. View Article : Google Scholar : PubMed/NCBI
22	Bruckdorfer R: The basics about nitric oxide. Mol Aspects Med. 26:3–31. 2005. View Article : Google Scholar : PubMed/NCBI
23	Madhusoodanan KS and Murad F: NO-cGMP signaling and regenerative medicine involving stem cells. Neurochem Res. 32:681–694. 2007. View Article : Google Scholar : PubMed/NCBI
24	Russo I, Viretto M, Doronzo G, et al: A short-term incubation with high glucose impairs VASP phosphorylation at serine 239 in response to the nitric oxide/cGMP pathway in vascular smooth muscle cells: role of oxidative stress. Biomed Res Int. 2014:3289592014. View Article : Google Scholar : PubMed/NCBI
25	Lambrechts A, Kwiatkowski AV, Lanier LM, et al: cAMP-dependent protein kinase phosphorylation of EVL, a Mena/VASP relative, regulates its interaction with actin and SH3 domains. J Biol Chem. 275:36143–36151. 2000. View Article : Google Scholar : PubMed/NCBI
26	Butt E, Abel K, Krieger M, et al: cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. J Biol Chem. 269:14509–14517. 1994.PubMed/NCBI
27	Tao Y, Chen YC, Sang JR and Xu WR: Phosphorylation of vasodilator stimulated phosphoprotein is correlated with cell cycle progression in HeLa cells. Mol Med Rep. 3:657–662. 2010.PubMed/NCBI
28	Roux PP and Blenis J: ERK and p38 MAPK-activated protein kinases: a family of protein kinases with diverse biological functions. Microbiol Mol Biol Rev. 68:320–344. 2004. View Article : Google Scholar : PubMed/NCBI
29	Sang JR, Chen YC and Tao Y: Nitric oxide inhibits gastric cancer cell growth through the modulation of the Akt pathway. Mol Med Rep. 4:1163–1167. 2011.PubMed/NCBI