IL‑6 and IL‑8 enhance factor H binding to the cell membranes

Popek,Sylwia; Kapka‑Skrzypczak,Lucyna; Sawicki,Krzysztof; Wolińska,Ewa; Skrzypczak,Maciej; Czajka,Magdalena

doi:10.3892/mmr.2016.5012

IL‑6 and IL‑8 enhance factor H binding to the cell membranes

Authors:
- Sylwia Popek
- Lucyna Kapka‑Skrzypczak
- Krzysztof Sawicki
- Ewa Wolińska
- Maciej Skrzypczak
- Magdalena Czajka
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Affiliations: Department of Molecular Biology and Translational Research, Institute of Rural Health, 20‑090 Lublin, Poland, Department of Medical Biology and Translational Research, Faculty of Medicine, University of Information Technology and Management, 35‑225 Rzeszow, Poland, Department of Pathology, Medical University of Warsaw, 02‑091 Warsaw, Poland, Second Department of Gynecology, Medical University of Lublin, 20‑954 Lublin, Poland
Published online on: March 18, 2016 https://doi.org/10.3892/mmr.2016.5012
Pages: 3886-3894
Copyright: © Popek et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to assess the role of interleukin (IL)‑6 and IL‑8 on the expression of fluid‑phase complement inhibitor, factor H (FH), and FH‑like protein 1 (FHL‑1), in the A2780 ovarian carcinoma cell line. This cell line does not normally produce IL‑6, however, is IL‑6 responsive due to the presence of receptor for IL‑6. The presence of FH and FHL‑1 in the cell lysates was confirmed by western blotting. The levels of FH and FHL‑1 in the medium were determined by enzyme‑linked immunosorbent assay. To evaluate gene expression, reverse transcription‑quantitative polymerase chain reaction was performed. The cellular localization of FH and FHL‑1 in ovarian cancer cells was assessed by immunofluorescence. The present study revealed that FH, contrary to FHL‑1, was secreted by ovarian cancer cells, however, this process was independent of IL stimulation. No significant differences were observed in the concentration of FH in the control cells, when compared with the samples treated with IL‑6/IL‑8. The results of western blotting revealed that the protein expression levels of FH and FHL‑1 were not regulated by IL‑6 and IL‑8 in a dose‑dependent manner. Immunofluorescence analysis confirmed that the A2780 ovarian cancer cell line expressed both membrane bound and intracellular forms of FH and FHL‑1. The present data revealed that the A2780 cells expressed and secreted FH protein and are also able to bind FH and FHL‑1. This may influence the efficiency of complement mediated immunotherapy.

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1	Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J, Rosso S, Coebergh JW, Comber H, Forman D and Bray F: Cancer incidence and mortality patterns in Europe: Estimates for 40 countries in 2012. Eur J Cancer. 49:1374–1403. 2013. View Article : Google Scholar : PubMed/NCBI
2	Wcislo G and Szczylik C: Foreword. Ovarian cancer-pathobiology, diagnosis and overview of contemporary methods of treatment. Termedia; Poznan: pp. 9–10. 2010
3	Urban A and Miszczyk L: Ovarian cancer-diagnostical and therapeutical dilema in oncological gynecology. Wspolczesna Onkol. 7:294–300. 2003.In Polish.
4	Gancz D and Fishelson Z: Cancer resistance to complement-dependent cytotoxicity (CDC): Problem-oriented research and development. Mol Immunol. 46:2794–2800. 2009. View Article : Google Scholar : PubMed/NCBI
5	Sarma JV and Ward PA: The complement system. Cell Tissue Res. 343:227–235. 2011. View Article : Google Scholar :
6	Fishelson Z, Donin N, Zell S, Schultz S and Kirschfink M: Obstacles to cancer immunotherapy: Expression of membrane complement regulatory proteins (mCRPs) in tumors. Mol Immunol. 40:109–123. 2003. View Article : Google Scholar : PubMed/NCBI
7	Liu B, Nash J, Runowicz C, Swede H, Stevens R and Li Z: Ovarian cancer immunotherapy: Opportunities, progresses and challenges. J Hemato Oncol. 3(7)2010. View Article : Google Scholar
8	Junnikkala S, Hakulinen J, Jarva H, Manuelian T, Bjørge L, Bützow R, Zipfel PF and Meri S: Secretion of soluble complement inhibitors factor H and factor H-like protein 1 (FHL-1) by ovarian tumor cells. Br J Cancer. 87:1119–1127. 2002. View Article : Google Scholar : PubMed/NCBI
9	Rodriguez de Cordoba S, Esparza-Gordillo J, Goicoechea de Jorge E, Lopez-Trascasa M and Sánchez-Corral P: The human complement factor H: Functional roles, genetic variations and disease associations. Mol Immunol. 41:355–367. 2004. View Article : Google Scholar : PubMed/NCBI
10	Kryczek I, Gryboś M and Lange A: Biological and clinical impact of IL-6 production by ovarian carcinoma cells. Wspolczesna Onkol. 3:195–198. 1999.In Polish.
11	Nowak M, Głowacka E, Szpakowski M, Szyllo K, Malinowski A, Kulig A, Tchórzewski H and Wilczyński J: Proinflamatory and immunosupresive serum, ascites and cyst fluid cytokines in patients with early and advenced ovarian cancer and benign ovarian tumors. Neuro Endocrinol Lett. 31:375–383. 2010.
12	Chou CH, Wei LH, Kuo ML, Huang YJ, Lai KP, Chen CA and Hsieh CY: Up-regulation of interleukin-6 in human ovarian cancer cell via a Gi/PI3K-Akt/NF-kappaB pathway by lysophosphatidic acid, an ovarian cancer-activating factor. Cancerogenesis. 26:45–52. 2005. View Article : Google Scholar
13	Dijkgraaf EM, Welters MJ, Nortier JW, van der Burg SH and Kroep JR: Interleukin-6/interleukin-6 receptor pathway as a new therapy target in epithelial ovarian cancer. Curr Pharm Des. 18:3816–3827. 2012. View Article : Google Scholar : PubMed/NCBI
14	Dobrzycka B, Mackowiak-Matejczyk B, Terlikowska KM, Kulesza-Bronczyk B, Kinalski M and Terlikowski SJ: Serum levels of IL-6, IL-8 and CRP as prognostic factors in epithelial ovarian cancer. Eur Cytokine Netw. 24:106–113. 2013.PubMed/NCBI
15	Wang Y, Niu XL, Qu Y, Wu J, Zhu YQ, Sun WJ and Li LZ: Autocrine production of interleukin-6 confers cisplatin and paclitaxel resistance in ovarian cancer cells. Cancer Lett. 295:110–123. 2010. View Article : Google Scholar : PubMed/NCBI
16	Spiller OB, Criado-García O, Rodríguez De Córdoba S and Morgan BP: Cytokine-mediated up-regulation of CD55 and CD59 protects human hepatoma cells from cells from complement attack. Clin Exp Immunol. 121:234–241. 2000. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
18	Ajona D, Castaño Z, Garayoa M, Zudaire E, Pajares MJ, Martinez A, Cuttitta F, Montuenga LM and Pio R: Expression of complement factor H by lung cancer cells: Effects on the activation of the alternative pathway of complement. Cancer Res. 64:6310–6318. 2004. View Article : Google Scholar : PubMed/NCBI
19	Bjørge L, Hakulinen J, Vintermy OK, Jarva H, Jensen TS, Iversen OE and Meri S: Ascitic complement system in ovarian cancer. Br J Cancer. 92:895–905. 2005. View Article : Google Scholar : PubMed/NCBI
20	Wang Y, Yang J, Gao Y, Du Y, Bao L, Niu W and Yao Z: Regulatory effect of E2, IL-6 and IL-8 on the Growth of epithelial ovarian cancer cells. Cell Mol Immunol. 2:365–372. 2005.PubMed/NCBI
21	Józsi M and Zipfel PF: Factor H family proteins and human diseases. Trends Immunol. 29:380–387. 2008. View Article : Google Scholar : PubMed/NCBI
22	Kapka-Skrzypczak L, Wolinska E, Szparecki G, Wilczynski GM, Czajka M and Skrzypczak M: CD55, CD59, factor H and factor H-like 1 gene expression analysis in tumors of the ovary and corpus uteri origin. Immunol Lett. 167:67–71. 2015. View Article : Google Scholar : PubMed/NCBI
23	Thibault B, Castells M, Delord JP and Couderc B: Ovarian cancer microenvironment: Implications for cancer dissemination and chemoresistance acquisition. Cancer Metastasis Rev. 33:17–39. 2014. View Article : Google Scholar
24	Józsi M, Manuelian T, Heinen S, Oppermann M and Zipfel PF: Attachment of the soluble complement regulator factor H to cell and tissue surfaces: Relevance for pathology. Histol Histopathol. 19:251–258. 2004.PubMed/NCBI
25	Kopp A, Hebecker M, Svobodová E, Józsi M and Factor H: A complement regulator in health and disease, and a mediator of cellular interactions. Biomolecules. 2:46–75. 2012. View Article : Google Scholar : PubMed/NCBI