title: IL‑6 and IL‑8 enhance factor H binding to the cell membranes.
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.
authors.affiliation: Department of Molecular Biology and Translational Research, Institute of Rural Health, 20‑090 Lublin, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Popek
authors.affiliation: Department of Medical Biology and Translational Research, Faculty of Medicine, University of Information Technology and Management, 35‑225 Rzeszow, Poland.
authors.forename: Lucyna
authors.initials: L
authors.lastname: Kapka-Skrzypczak
authors.affiliation: Department of Molecular Biology and Translational Research, Institute of Rural Health, 20‑090 Lublin, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Sawicki
authors.affiliation: Department of Pathology, Medical University of Warsaw, 02‑091 Warsaw, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Wolińska
authors.affiliation: Second Department of Gynecology, Medical University of Lublin, 20‑954 Lublin, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Skrzypczak
authors.affiliation: Department of Molecular Biology and Translational Research, Institute of Rural Health, 20‑090 Lublin, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Czajka
mesh_terms: D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism* / Q000473:pathology; D017242:Complement Factor H / Q000378:metabolism*; D005260:Female; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism*; D016209:Interleukin-8 / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D060588:LIM Domain Proteins / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000473:pathology; D011485:Protein Binding
chemical_list: C508606:CXCL8 protein, human; C101149:FHL1 protein, human; C508600:IL6 protein, human; D015850:Interleukin-6; D016209:Interleukin-8; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D009124:Muscle Proteins; D009363:Neoplasm Proteins; D017242:Complement Factor H