Increased expression of p21Waf1/Cip1 and JNK with costimulation of prostate cancer cell activation by an siRNA Egr-1 inhibitor

Parra,Eduardo; Gutiérrez,Luis; Ferreira,Jorge

doi:10.3892/or.2013.2503

Increased expression of p21Waf1/Cip1 and JNK with costimulation of prostate cancer cell activation by an siRNA Egr-1 inhibitor

Authors:
- Eduardo Parra
- Luis Gutiérrez
- Jorge Ferreira
View Affiliations

Affiliations: Laboratory of Experimental Biomedicine, University of Tarapaca, Campus Esmeralda, Iquique, Chile, Faculty of Sciences, Arturo Prat University, Iquique, Chile, Programme of Molecular and Clinical Pharmacology, ICBM, Medical Faculty, University of Chile, Santiago, Chile
Published online on: May 28, 2013 https://doi.org/10.3892/or.2013.2503
Pages: 911-916

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

The p21Waf1/Cip1 protein (hereafter, p21) and the c‑Jun N-terminal kinase (JNK) are two well-characterized cell modulators that play a crucial role in cell differentiation, senescence and apoptosis. Here, we report that transcription of the p21Waf1/Cip1 and JNK-1 genes is affected by inhibition of the early growth response-1 (Egr-1) in response to a small interfering RNA [siRNA)-Egr-1] in LNCaP and PC-3 prostate carcinoma cell lines. The expression levels of protein were determined by western blotting, and apoptosis was measured by propidium iodide staining and flow cytometric analysis. Inhibition of Egr-1, p21 and JNK-1 was carried out by siRNAs. LNCaP and PC-3 cells exhibited readily detectable Egr-1, JNK and p21, even in low serum medium without the addition of other exogenous agents. The expression of Egr-1, p21 and JNK was strongly increased after treatment of the cells with TPA, tumor necrosis factor-α (TNF-α) or arsenite. Suppression of Egr-1 expression by siRNA abrogated the ability of TPA to induce Egr-1 and JNK-1 activities, moderately increasing the p21 activity and abrogating the anti-apoptotic effect of Egr-1 observed in the prostate cancer cell lines. Moreover, blockade of p21 and JNK was unable to decrease the activity of Egr-1, while siRNA against p21 abrogated the pro‑apoptotic effect of p21. The results demonstrated that Egr-1 acts as a key player in prostate tumor cell growth and survival, while p21 plays a key pro‑apoptotic role in LNCaP and PC-3 prostate carcinoma cell lines.

View Figures

View References

1	Stewart AZ, Leach DS and Pietenpol AJ: p21(Waf1/Cip1) inhibition of cyclin E/Cdk2 activity prevents endoreduplication after mitotic spindle disruption. Mol Cell Biol. 19:205–215. 1999.PubMed/NCBI
2	Satyanarayana A, Hilton MB and Kaldis P: p21 inhibits Cdk1 in the absence of Cdk2 to maintain the G1/S phase DNA damage checkpoint. Mol Biol Cell. 19:65–77. 2008. View Article : Google Scholar : PubMed/NCBI
3	Heasley LE and Han SY: JNK regulation of oncogenesis. Mol Cells. 21:167–173. 2006.
4	Matsukawa J, Matsuzawa A, Takeda K and Ichijo H: The ASK1-MAP kinase cascades in mammalian stress response. J Biochem. 136:261–265. 2004. View Article : Google Scholar : PubMed/NCBI
5	Potapova O, Haghighi A, Bost F, Liu C, Birrer M, Gjerset R and Mercola D: The Jun kinase/stress-activated protein kinase pathway functions to regulate DNA repair and inhibition of the pathway sensitizes tumor cells to cisplatin. J Biol Chem. 272:14041–14044. 1997. View Article : Google Scholar : PubMed/NCBI
6	Gupta S, Campbell D, Dérijard B and Davis RJ: Transcription factor ATF2 regulation by the JNK signal transduction pathway. Science. 267:389–393. 1995. View Article : Google Scholar : PubMed/NCBI
7	Bocco JL, Bahr A, Goetz J, Hauss C, Kallunki T, Kedinger C and Chatton B: In vivo association of ATFa with JNK/SAP kinase activities. Oncogene. 12:1971–1980. 1996.PubMed/NCBI
8	Hibi M, Lin A, Smeal T, Minden A and Karin M: Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 7:2135–2148. 1993. View Article : Google Scholar : PubMed/NCBI
9	Janknecht R and Hunter T: Convergence of MAP kinase pathways on the ternary complex factor Sap-1a. EMBO J. 16:1620–1627. 1997. View Article : Google Scholar : PubMed/NCBI
10	Dhanasekaran DN and Reddy EP: JNK signaling in apoptosis. Oncogene. 27:6245–6251. 2008. View Article : Google Scholar : PubMed/NCBI
11	Liu J and Lin A: Role of JNK activation in apoptosis: a double-edged sword. Cell Res. 15:36–42. 2005. View Article : Google Scholar : PubMed/NCBI
12	Parra E and Ferreira J: Knockdown of the c-Jun-N-terminal kinase expression by siRNA inhibits MCF-7 breast carcinoma cell lines growth. Oncol Rep. 24:1339–1345. 2010. View Article : Google Scholar : PubMed/NCBI
13	Parra E: Activation of MAP kinase family members triggered by TPA or ionomycin occurs via the protein phosphatase 4 pathway in Jurkat leukemia T cells. Mol Med Rep. 5:773–778. 2012.PubMed/NCBI
14	Parra E: Inhibition of JNK-1 by small interference RNA induces apoptotic signalling in PC-3 prostate cancer cells. Int J Mol Med. 30:923–930. 2012.PubMed/NCBI
15	Beckmann AM and Wilce PA: EGR transcription factors in the nervous system. Neurochem Int. 31:477–510. 1997. View Article : Google Scholar
16	Whitlock NC, Bahn JH, Lee SH, Eling TE and Baek SJ: Resveratrol-induced apoptosis is mediated by early growth response-1, Krüppel-like factor 4, and activating transcription factor 3. Cancer Prev Res. 4:116–127. 2011.PubMed/NCBI
17	Eichelberger L, Koch MO, Eble JN, Ulbright TM, Juliar BE and Cheng L: Maximum tumor diameter is an independent predictor of prostate-specific antigen recurrence in prostate cancer. Mod Pathol. 18:886–890. 2005. View Article : Google Scholar : PubMed/NCBI
18	Houston P, Campbell CJ, Svaren J, Milbrandt J and Braddock M: The transcriptional corepressor NAB2 blocks EGR-1-mediated growth factor activation and agiogenesis. Biochem Biophys Res Commun. 283:480–486. 2001. View Article : Google Scholar : PubMed/NCBI
19	Collins S, Lutz MA, Zank PE, Anders RA, Kersh GJ and Powell JD: Opposing regulation of T cell function by EGR-1/NAB2 and EGR-2/EGR-3. Eur J Immunol. 38:528–536. 2008. View Article : Google Scholar : PubMed/NCBI
20	Parra E, Ortega A and Saenz L: Down-regulation of EGR-1 by siRNA inhibits growth of human prostate carcinoma cell line PC-3. Oncol Rep. 22:1513–1518. 2009.PubMed/NCBI
21	Pignatelli M, Luna-Medina R, Pérez-Rendón A, Santos A and Perez-Castillo A: The transcription factor early growth response factor-1 (EGR-1) promotes apoptosis of neuroblastoma cells. Biochem J. 373:739–746. 2003. View Article : Google Scholar : PubMed/NCBI
22	Banks MF, Gerasimovskaya EV, Tucker DA, Frid MG, Carpenter TC and Stenmark KR: EGR-1 antisense oligonucleotides inhibit hypoxia-induced proliferation of pulmonary artery adventitial fibroblasts. J Appl Physiol. 98:732–738. 2005. View Article : Google Scholar
23	Sperandio S, Fortin J, Sasik R, Robitaille L, Corbeil J and de Belle I: The transcription factor EGR1 regulates the HIF-1alpha gene during hypoxia. Mol Carcinog. 48:38–44. 2009. View Article : Google Scholar : PubMed/NCBI
24	Parra E and Ferreira J: The Effect of siRNA-EGR-1 and camptothecin on growth and chemosensitivity of breast cancer cell lines. Oncol Rep. 22:1159–1165. 2010.PubMed/NCBI
25	Adamson ED and Mercola D: EGR1 transcription factor: multiple roles in prostate tumor cell growth and survival. Tumour Biol. 23:93–102. 2002. View Article : Google Scholar : PubMed/NCBI
26	Catania MV, Copani A, Calogero A, Ragonese GI, Condorelli DF and Nicoletti F: An enhanced expression of the immediate early gene, EGR-1, is associated with neuronal apoptosis in culture. Neuroscience. 91:1529–1538. 1999. View Article : Google Scholar : PubMed/NCBI
27	Nair P, Muthukkumar S, Sells SF, Han S, Sukhatme VP and Rangnekar VM: Early growth response-1-dependent apoptosis is mediated by p53. J Biol Chem. 272:20131–20138. 1997. View Article : Google Scholar : PubMed/NCBI
28	Calogero A, Arcella A, De Gregorio G, Porcellini A, Mercola D, Liu C, Lombari V, Zani M, Giannini G, et al: The early growth response gene EGR-1 behaves as a suppressor gene that is down-regulated independent of ARF/Mdm2 but not p53 alterations in fresh human gliomas. Clin Cancer Res. 7:2788–2796. 2001.PubMed/NCBI
29	Stevenson M: Therapeutic potential of RNA interference. N Engl J Med. 351:1772–1777. 2004. View Article : Google Scholar : PubMed/NCBI
30	Sui G, Soohoo C, Affar el B, Gay F and Shi Y, Forrester WC and Shi Y: A DNA vector-based RNAi technology to suppress gene expression in mammalian cells. Proc Natl Acad Sci USA. 99:5515–5520. 2002. View Article : Google Scholar : PubMed/NCBI
31	Parra E, Ferreira J and Saenz L: Inhibition of Egr-1 by siRNA in prostate carcinoma cell lines is associated with decreased expression of AP-1 and NF-κB. Int J Mol Med. 28:847–853. 2011.PubMed/NCBI
32	Ciccarelli C, Marampon F, Scoglio A, Mauro A, Giacinti C, De Cesaris P and Zani BM: p21^WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells. Mol Cancer. 4:412005. View Article : Google Scholar
33	Todd DE, Densham RM, Molton SA, Balmanno K, Newson C, Weston CR, Garner AP, Scott L and Cook SJ: ERK1/2 and p38 cooperate to induce a p21^CIP1-dependent G1 cell cycle arrest. Oncogene. 23:3284–3295. 2004. View Article : Google Scholar : PubMed/NCBI
34	Ragione FD, Cucciolla V, Criniti V, Indaco S, Borriello A and Zappia V: p21^Cip1 gene expression is modulated by EGR1: a novel regulatory mechanism involved in the resveratrol antiproliferative effect. J Bio Chem. 278:23360–23368. 2003.
35	Boutros T, Chevet E and Metrakos P: Mitogen-activated protein (MAP) kinase/MAP kinase phosphatase regulation: roles in cell growth, death, and cancer. Pharmacol Rev. 60:261–310. 2008. View Article : Google Scholar : PubMed/NCBI
36	Kim MJ, Choi SY, Park IC, Hwang SG, Kim C, Choi YH, Kim H, Lee KH and Lee SJ: Opposing roles of c-Jun NH2-terminal kinase and p38 mitogen-activated protein kinase in the cellular response to ionizing radiation in human cervical cancer cells. Mol Cancer Res. 6:1718–1731. 2008. View Article : Google Scholar
37	Wu J, Sun J and Xue Y: Involvement of JNK and P53 activation in G2/M cell cycle arrest and apoptosis induced by titanium dioxide nanoparticles in neuron cells. Toxicol Lett. 199:269–276. 2010. View Article : Google Scholar : PubMed/NCBI
38	Choi BH, Kim CG, Bae YS, Lim Y, Lee YH and Shin SY: p21^Waf1/Cip1 expression by curcumin in U-87MG human glioma cells: role of early growth response-1 expression. Cancer Res. 68:1369–1377. 2008.
39	Clark JA, Black AR, Leontieva OV, Frey MR, Pysz MA, Kunneva L, Woloszynska-Read A, Roy D and Black JD: Involvement of the ERK signaling cascade in protein kinase C-mediated cell cycle arrest in intestinal epithelial cells. J Biol Chem. 279:9233–9247. 2004. View Article : Google Scholar : PubMed/NCBI