S100B-p53 disengagement by pentamidine promotes apoptosis and inhibits cellular migration via aquaporin-4 and metalloproteinase-2 inhibition in C6 glioma cells

Capoccia,Elena; Cirillo,Carla; Marchetto,Annalisa; Tiberi,Samanta; Sawikr,Youssef; Pesce,Marcella; D'Alessandro,Alessandra; Scuderi,Caterina; Sarnelli,Giovanni; Cuomo,Rosario; Steardo,Luca; Esposito,Giuseppe

doi:10.3892/ol.2015.3091

S100B-p53 disengagement by pentamidine promotes apoptosis and inhibits cellular migration via aquaporin-4 and metalloproteinase-2 inhibition in C6 glioma cells

Authors:
- Elena Capoccia
- Carla Cirillo
- Annalisa Marchetto
- Samanta Tiberi
- Youssef Sawikr
- Marcella Pesce
- Alessandra D'Alessandro
- Caterina Scuderi
- Giovanni Sarnelli
- Rosario Cuomo
- Luca Steardo
- Giuseppe Esposito
View Affiliations

Affiliations: Department of Physiology and Pharmacology ̔Vittorio Erspamer̓, Sapienza University of Rome, Rome I‑00185, Italy, Laboratory for Enteric Neuroscience, Translational Research Center for Gastrointestinal Disorders, University of Leuven, Leuven 3000, Belgium, Department of Clinical and Experimental Medicine, Gastroenterology Unit, University of Naples Federico II, Naples I‑80131, Italy
Published online on: April 1, 2015 https://doi.org/10.3892/ol.2015.3091
Pages: 2864-2870

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

S100 calcium-binding protein B (S100B) is highly expressed in glioma cells and promotes cancer cell survival via inhibition of the p53 protein. In melanoma cells, this S100B‑p53 interaction is known to be inhibited by pentamidine isethionate, an antiprotozoal agent. Thus, the aim of the present study was to evaluate the effect of pentamidine on rat C6 glioma cell proliferation, migration and apoptosis in vitro. The change in C6 cell proliferation following treatment with pentamidine was determined by performing a 3‑[4,5‑dimethylthiazol‑2‑yl]‑2,5 diphenyltetrazolium bromide‑formazan assay. Significant dose‑dependent decreases in proliferation were observed at pentamidine concentrations of 0.05 µM (58.5±5%; P<0.05), 0.5 µM (40.6±7%; P<0.01) and 5 µM (13±4%; P<0.001) compared with the control (100% viability). Furthermore, treatment with 0.05, 0.5 and 5 µM pentamidine was associated with a significant increase in apoptosis versus the untreated cells, as determined by DNA fragmentation assays, immunofluorescence analysis of C6 chromatin using Hoechst staining, and immunoblot analysis of B‑cell lymphoma‑2 (Bcl‑2)‑associated X protein (100%, P<0.05; 453%, P<0.01; and 1000%, P<0.001, respectively) and Bcl‑2 (‑60%, P<0.001; ‑80.13%, P<0.001; ‑95%, P<0.001, respectively). In addition, the administration of 0.05, 0.5 and 5 µM pentamidine significantly upregulated the protein expression levels of p53 (681±87.5%, P<0.05; 1244±94.3%, P<0.01; and 2244±111%, P<0.001, respectively), and significantly downregulated the expression levels of matrix metalloproteinase‑2 (42±2.3%, P<0.05; 71±2.5%, P<0.01; and 95.8±3.3%, P<0.001, respectively) and aquaporin 4 (38±2.5%, P<0.05; 69±2.6%, P<0.01; and 88±3.0%, P<0.001, respectively), compared with the untreated cells. The wound healing assay demonstrated that cell migration was significantly impaired by treatment with 0.05, 0.5 and 5 µM pentamidine compared with untreated cells (88±4.2%, P<0.05; 64±2%, P<0.01; and 42±3.1%, P<0.001, respectively). Although additional in vivo studies are required to clarify the current in vitro data, the present study indicates that pentamidine and S100B‑p53 inhibitors may represent a novel approach for the treatment of glioma.

View Figures

View References

1	Mamelak AN and Jacoby DB: Targeted delivery of antitumoral therapy to glioma and other malignancies with synthetic chlorotoxin (TM-601). Expert Opin Drug Deliv. 4:175–186. 2007. View Article : Google Scholar : PubMed/NCBI
2	Goodenberger ML and Jenkins RB: Genetics of adult glioma. Cancer Genet. 205:613–621. 2012. View Article : Google Scholar : PubMed/NCBI
3	Crocetti E, Trama A, Stiller C, Caldarella A, Soffietti R, Jaal J, et al: RARECARE Working Group: Epidemiology of glial and non-glial brain tumours in Europe. Eur J Cancer. 48:1532–1542. 2012. View Article : Google Scholar : PubMed/NCBI
4	Stupp R, Pavlidis N and Jelic S: ESMO Guidelines Task Force: ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of malignant glioma. Ann Oncol. 16:(Suppl 1). i64–i65. 2005. View Article : Google Scholar : PubMed/NCBI
5	Maher EA, Furnari FB, Bachoo RM, Rowitch DH, Louis DN, Cavenee WK and DePinho RA: Malignant glioma: genetics and biology of a grave matter. Genes Dev. 15:1311–1333. 2001. View Article : Google Scholar : PubMed/NCBI
6	Giese A, Rief MD, Loo MA and Berens ME: Determinants of human astrocytoma migration. Cancer Res. 54:3897–3904. 1994.PubMed/NCBI
7	Louis DN, Pomeroy SL and Cairncross JG: Focus on central nervous system neoplasia. Cancer Cell. 1:125–128. 2002. View Article : Google Scholar : PubMed/NCBI
8	Donato R, Sorci G, Riuzzi F, et al: S100B's double life: intracellular regulator and extracellular signal. Biochim Biophys Acta. 1793:1008–1022. 2009. View Article : Google Scholar : PubMed/NCBI
9	Chaves ML, Camozzato AL, Ferreira ED, et al: Serum levels of S100B and NSE proteins in Alzheimer's disease patients. J Neuroinflammation. 7:62010. View Article : Google Scholar : PubMed/NCBI
10	Sathe K, Maetzler W, Lang JD, et al: S100B is increased in Parkinson's disease and ablation protects against MPTP-induced toxicity through the RAGE and TNF-α pathway. Brain. 135:3336–3347. 2012. View Article : Google Scholar : PubMed/NCBI
11	Weide B, Richter S, Büttner P, et al: Serum S100B, lactate dehydrogenase and brain metastasis are prognostic factors in patients with distant melanoma metastasis and systemic therapy. PLoS One. 8:e816242013. View Article : Google Scholar : PubMed/NCBI
12	Lin J, Yang Q, Yan Z, Markowitz J, Wilder PT, Carrier F and Weber DJ: Inhibiting S100B restores p53 levels in primary malignant melanoma cancer cells. J Biol Chem. 279:34071–34077. 2004. View Article : Google Scholar : PubMed/NCBI
13	Rustandi RR, Baldisseri DM and Weber DJ: Structure of the negative regulatory domain of p53 bound to S100B(betabeta). Nat Struct Biol. 7:570–574. 2000. View Article : Google Scholar : PubMed/NCBI
14	Millward TA, Heizmann CW, Schäfer BW and Hemmings BA: Calcium regulation of Ndr protein kinase mediated by S100 calcium-binding proteins. EMBO J. 17:5913–5922. 1998. View Article : Google Scholar : PubMed/NCBI
15	Arcuri C, Bianchi R, Brozzi F and Donato R: S100B increases proliferation in PC12 neuronal cells and reduces their responsiveness to nerve growth factor via Akt activation. J Biol Chem. 280:4402–4414. 2005. View Article : Google Scholar : PubMed/NCBI
16	Hartman KG, McKnight LE, Liriano MA and Weber DJ: The evolution of S100B inhibitors for the treatment of malignant melanoma. Future Med Chem. 5:97–109. 2013. View Article : Google Scholar : PubMed/NCBI
17	Smith J, Stewart BJ, Glaysher S, Peregrin K, Knight LA, Weber DJ and Cree IA: The effect of pentamidine on melanoma ex vivo. Anticancer Drugs. 21:181–185. 2010. View Article : Google Scholar : PubMed/NCBI
18	Sibenaller ZA, Etame AB, Ali MM, Barua M, Braun TA, Casavant TL and Ryken TC: Genetic characterization of commonly used glioma cell lines in the rat animal model system. Neurosurg Focus. 19:E12005. View Article : Google Scholar : PubMed/NCBI
19	Amberger VR, Hensel T, Ogata N and Schwab ME: Spreading and migration of human glioma and rat C6 cells on central nervous system myelin in vitro is correlated with tumor malignancy and involves a metalloproteolytic activity. Cancer Res. 58:149–158. 1998.PubMed/NCBI
20	Gunnersen JM, Spirkoska V, Smith PE, Danks RA and Tan SS: Growth and migration markers of rat C6 glioma cells identified by serial analysis of gene expression. Glia. 32:146–154. 2000. View Article : Google Scholar : PubMed/NCBI
21	Grobben B, DE Deyn PP and Slegers H: Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion. Cell Tissue Res. 310:257–270. 2002. View Article : Google Scholar : PubMed/NCBI
22	Schlegel J, Piontek G, Kersting M, Schuermann M, Kappler R, Scherthan H, et al: The p16/Cdkn2a/Ink4a gene is frequently deleted in nitrosourea-induced rat glial tumors. Pathobiology. 67:202–206. 1999. View Article : Google Scholar : PubMed/NCBI
23	Asai A, Miyagi Y, Hashimoto H, Lee SH, Mishima K, Sugiyama A, et al: Modulation of tumor immunogenicity of rat glioma cells by s-Myc expression: eradication of rat gliomas in vivo. Cell Growth Differ. 11:1153–1158. 1994.
24	Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
25	Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72:248–254. 1976. View Article : Google Scholar : PubMed/NCBI
26	Renault-Mihara F, Beuvon F, Iturrioz X, Canton B, De Bouard S, Léonard N, et al: Phosphoprotein enriched in astrocytes-15 kDa expression inhibits astrocyte migration by a protein kinase C delta-dependent mechanism. Mol Biol Cell. 17:5141–5152. 2006. View Article : Google Scholar : PubMed/NCBI
27	Clarke J, Butowski N and Chang S: Recent advances in therapy for glioblastoma. Arch Neurol. 67:279–283. 2010. View Article : Google Scholar : PubMed/NCBI
28	Rieger L, Weller M, Bornemann A, Schabet M, Dichgans J and Meyermann R: BCL-2 family protein expression in human malignant glioma: a clinical-pathological correlative study. J Neurol Sci. 155:68–75. 1998. View Article : Google Scholar : PubMed/NCBI
29	Paul-Samojedny M, Kokocińska D, Samojedny A, Mazurek U, Partyka R, Lorenz Z and Wilczok T: Expression of cell survival/death genes: Bcl-2 and Bax at the rate of colon cancer prognosis. Biochim Biophys Acta. 1741:25–29. 2005. View Article : Google Scholar : PubMed/NCBI
30	Pathak MK, Dhawan D, Lindner DJ, Borden EC, Farver C and Yi T: Pentamidine is an inhibitor of PRL phosphatases with anticancer activity. Mol Cancer Ther. 1:1255–1264. 2002.PubMed/NCBI
31	Zimmer DB, Lapidus RG and Weber DJ: In vivo screening of S100B inhibitors for melanoma therapy. Methods Mol Biol. 963:303–317. 2013. View Article : Google Scholar : PubMed/NCBI
32	Du R, Petritsch C, Lu K, Liu P, Haller A, Ganss R, et al: Matrix metalloproteinase-2 regulates vascular patterning and growth affecting tumor cell survival and invasion in GBM. Neuro Oncol. 10:254–264. 2008. View Article : Google Scholar : PubMed/NCBI
33	Ding T, Zhou Y, Sun K, Jiang W, Li W, Liu X, et al: Knockdown a water channel protein, aquaporin-4, induced glioblastoma cell apoptosis. PLoS One. 8:e667512013. View Article : Google Scholar : PubMed/NCBI
34	Verkman AS: Aquaporins in clinical medicine. Annu Rev Med. 63:303–316. 2012. View Article : Google Scholar : PubMed/NCBI
35	Giese A: Glioma invasion-pattern of dissemination by mechanisms of invasion and surgical intervention, pattern of gene expression and its regulatory control by tumorsuppressor p53 and proto-oncogene ETS-1. Acta Neurochir Suppl. 88:153–162. 2003.PubMed/NCBI
36	Sanderson L, Dogruel M, Rodgers J, et al: Pentamidine movement across the murine blood-brain and blood-cerebrospinal fluid barriers: effect of trypanosome infection, combination therapy, P-glycoprotein, and multidrug resistance-associated protein. J Pharmacol Exp Ther. 329:967–977. 2009. View Article : Google Scholar : PubMed/NCBI
37	Kotthaus J, Kotthaus J, Schade D, et al: New prodrugs of the antiprotozoal drug pentamidine. ChemMedChem. 9:2233–2242. 2011. View Article : Google Scholar
38	Raseroka BH and Ormerod WE: The trypanocidal effect of drugs in different parts of the brain. Trans R Soc Trop Med Hyg. 80:634–641. 1986. View Article : Google Scholar : PubMed/NCBI