PRKACB is downregulated in non‑small cell lung cancer and exogenous PRKACB inhibits proliferation and invasion of LTEP‑A2 cells

Chen,Yong; Gao,Ying; Tian,Ye; Tian,Da‑Li

doi:10.3892/ol.2013.1294

PRKACB is downregulated in non‑small cell lung cancer and exogenous PRKACB inhibits proliferation and invasion of LTEP‑A2 cells

Authors:
- Yong Chen
- Ying Gao
- Ye Tian
- Da‑Li Tian
View Affiliations

Affiliations: Department of Thoracic Surgery, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning 110032, P.R. China, Department of Pathology, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning 110032, P.R. China
Published online on: April 8, 2013 https://doi.org/10.3892/ol.2013.1294
Pages: 1803-1808

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

Protein kinase cAMP‑dependent catalytic β (PRKACB) is a member of the Ser/Thr protein kinase family and a key effector of the cAMP/PKA‑induced signal transduction involved in numerous cellular process, including cell proliferation, apoptosis, gene transcription, metabolism and differentiation. In the present study, the expression pattern of PRKACB in non‑small cell lung cancer (NSCLC) and the effect of PRKACB upregulation on cell proliferation, apoptosis and invasion were investigated. PRKACB mRNA and protein expression was analyzed in the NSCLC tissue and corresponding normal tissues of 30 cases, using quantitative RT‑PCR and western blot analysis. A plasmid containing full‑length PRKACB was transfected into LTEP‑A2 cells to further investigate the effects of PRKACB overexpression on proliferation, apoptosis and invasion of the transfected cells, which were examined using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT), colony formation, flow cytometry and Transwell assays. The results revealed that the NSCLC tissues exhibited much lower levels of PRKACB mRNA and protein compared with their corresponding normal tissues. The upregulation of PRKACB decreased the numbers of proliferative, colony and invasive cells, while the apoptotic rates of transfected cells were increased. These data indicate that PRKACB is downregulated in NSCLC tissues and that upregulation of PRKACB may be an effective way to prevent the progression of NSCLC.

View Figures

View References

1	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	Molina JR, Yang P, Cassivi SD, Schild SE and Adjei AA: Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc. 83:584–594. 2008. View Article : Google Scholar : PubMed/NCBI
3	Skalhegg BS and Tasken K: Specificity in the cAMP/PKA signaling pathway: Differential expression, regulation, and subcellular localization of subunits of PKA. Front Biosci. 5:D678–D693. 2000. View Article : Google Scholar
4	Corbin JD, Sugden PH, West L, Flockhart DA, Lincoln TM and McCarthy D: Studies on the properties and mode of action of the purified regulatory subunit of bovine heart adenosine 3′:5′-monophosphate-dependent protein kinase. J Biol Chem. 253:3997–4003. 1978.
5	Cook SJ and McCormick F: Inhibition by cAMP of Ras-dependent activation of Raf. Science. 262:1069–1072. 1993. View Article : Google Scholar : PubMed/NCBI
6	Lang P, Gesbert F, Delespine-Carmagnat M, et al: Protein kinase A phosphorylation of RhoA mediates the morphological and functional effects of cyclic AMP in cytotoxic lymphocytes. EMBO J. 15:510–519. 1996.PubMed/NCBI
7	Schmitt JM and Stork PJ: PKA phosphorylation of Src mediates cAMP’s inhibition of cell growth via Rap1. Mol Cell. 9:85–94. 2002.
8	Michl P, Knobel B and Downward J: CUTL1 is phosphorylated by protein kinase A, modulating its effects on cell proliferation and motility. J Biol Chem. 281:15138–15144. 2006. View Article : Google Scholar : PubMed/NCBI
9	Larsen AC, Kvissel AK, Hafte TT, Avellan CI, Eikvar S, et al: Inactive forms of the catalytic subunit of protein kinase A are expressed in the brain of higher primates. FEBS J. 275:250–262. 2008. View Article : Google Scholar : PubMed/NCBI
10	Ørstavik S, Reinton N, Frengen E, Langeland BT, Jahnsen T and Skålhegg BS: Identification of novel splice variants of the human catalytic subunit Cbeta of cAMP-dependent protein kinase. Eur J Biochem. 268:5066–5073. 2001.PubMed/NCBI
11	Kvissel AK, Ørstavik S, Øistad P, Rootwelt T, Jahnsen T and Skålhegg BS: Induction of Cbeta splice variants and formation of novel forms of protein kinase A type II holoenzymes during retinoic acid-induced differentiation of human NT2 cells. Cell Signal. 16:577–587. 2004. View Article : Google Scholar
12	Kvissel AK, Ramberg H, Eide T, Svindland A, et al: Androgen dependent regulation of protein kinase A subunits in prostate cancer cells. Cell Signal. 19:401–409. 2007. View Article : Google Scholar : PubMed/NCBI
13	Wu KJ, Mattioli M, Morse HC III and Dalla-Favera R: c-MYC activates protein kinase A (PKA) by direct transcriptional activation of the PKA catalytic subunit beta (PKA-Cbeta) gene. Oncogene. 21:7872–7882. 2002. View Article : Google Scholar : PubMed/NCBI
14	Higuchi H, Yamashita T, Yoshikawa H and Tohyama M: PKA phosphorylates the p75 receptor and regulates its localization to lipid rafts. EMBO J. 22:1790–1800. 2003. View Article : Google Scholar : PubMed/NCBI
15	Schraders M, Jares P, Bea S, Schoenmakers EF, et al: Integrated genomic and expression profiling in mantle cell lymphoma: identification of gene-dosage regulated candidate genes. Br J Haematol. 143:210–221. 2008. View Article : Google Scholar : PubMed/NCBI
16	Erlbruch A, Hung CW, Seidler J, Borrmann K, et al: Uncoupling of bait-protein expression from the prey protein environment adds versatility for cell and tissue interaction proteomics and reveals a complex of CARP-1 and the PKA Cbeta1 subunit. Proteomics. 10:2890–2900. 2010. View Article : Google Scholar : PubMed/NCBI
17	Kaplan DR and Miller FD: Neurotrophin signal transduction in the nervous system. Curr Opin Neurobiol. 10:381–391. 2000. View Article : Google Scholar : PubMed/NCBI
18	Matsumoto H, Sakamoto A, Fujiwara M, et al: Cyclic AMP-mediated growth suppression and MAPK phosphorylation in thyroid papillary carcinoma cells. Mol Med Rep. 1:245–249. 2008.PubMed/NCBI
19	Nguyen GH, French R and Radhakrishna H: Protein kinase A inhibits lysophosphatidic acid induction of serum response factor via alterations in the actin cytoskeleton. Cell Signal. 16:1141–1151. 2004. View Article : Google Scholar : PubMed/NCBI
20	Chen TC, Hinton DR, Zidovetzki R and Hofman FM: Up-regulation of the cAMP/PKA pathway inhibits proliferation, induces differentiation, and leads to apoptosis in malignant gliomas. Lab Invest. 78:165–174. 1998.PubMed/NCBI
21	Cassoni P, Sapino A, Fortunati N, Munaron L, Chini B and Bussolati G: Oxytocin inhibits the proliferation of MDA-MB231 human breast-cancer cells via cyclic adenosine monophosphate and protein kinase A. Int J Cancer. 72:340–344. 1997. View Article : Google Scholar : PubMed/NCBI
22	Hewer RC, Sala-Newby GB, Wu YJ, Newby AC and Bond M: PKA and Epac synergistically inhibit smooth muscle cell proliferation. J Mol Cell Cardiol. 50:87–98. 2011. View Article : Google Scholar : PubMed/NCBI
23	Liu J, Li XD, Ora A, Heikkilä P, Vaheri A and Voutilainen R: cAMP-dependent protein kinase activation inhibits proliferation and enhances apoptotic effect of tumor necrosis factor-alpha in NCI-H295R adrenocortical cells. J Mol Endocrinol. 33:511–522. 2004. View Article : Google Scholar
24	D’Angelo G, Lee H and Weiner RI: cAMP-dependent protein kinase inhibits the mitogenic action of vascular endothelial growth factor and fibroblast growth factor in capillary endothelial cells by blocking Raf activation. J Cell Biochem. 67:353–366. 1997.
25	Hordijk PL, Verlaan I, Jalink K, van Corven EJ and Moolenaar WH: cAMP abrogates the p21ras-mitogen-activated protein kinase pathway in fibroblasts. J Biol Chem. 269:3534–3538. 1994.PubMed/NCBI
26	van Oirschot BA, Stahl M, Lens SM and Medema RH: Protein kinase A regulates expression of p27(kip1) and cyclin D3 to suppress proliferation of leukemic T cell lines. J Biol Chem. 276:33854–33860. 2001.PubMed/NCBI
27	Al-Wadei HA and Schuller HM: Cyclic adenosine monophosphate-dependent cell type-specific modulation of mitogenic signaling by retinoids in normal and neoplastic lung cells. Cancer Detect Prev. 30:403–411. 2006. View Article : Google Scholar : PubMed/NCBI