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TNF ‐ α ‐induced IL ‐6 and MMP ‐9 expression in immortalized ameloblastoma cell line established by hTERT
Author(s) -
Ohta K,
Naruse T,
Ishida Y,
Shigeishi H,
Nakagawa T,
Fukui A,
Nishi H,
Sasaki K,
Ogawa I,
Takechi M
Publication year - 2017
Publication title -
oral diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.953
H-Index - 87
eISSN - 1601-0825
pISSN - 1354-523X
DOI - 10.1111/odi.12594
Subject(s) - telomerase reverse transcriptase , telomerase , transfection , tumor necrosis factor alpha , cell culture , microbiology and biotechnology , biology , immortalised cell line , cancer research , immunology , gene , genetics
Objective Ameloblastoma ( AM ) shows locally invasive behaviour. However, biological investigations regarding regulation of gene expression associated with AM pathological features are difficult to perform, because AM cells can be passaged for a few generations due to senescence. We report a newly established immortalized AM cell line, AMB cells, by transfection with human telomerase reverse transcriptase ( hTERT ). Furthermore, we examined whether TNF ‐ α modulates bone resorption‐related genes, IL ‐6 and MMP ‐9 in cooperation with TGF ‐ β or IFN ‐ γ . Materials and methods Following transfection of an hTERT expression vector into AM cells using a non‐viral method, the effects of cytokines on the expressions of IL ‐6 and MMP ‐9 mRNA were examined using real‐time PCR . TNF ‐ α ‐induced NF ‐ κ B activity was examined by western blotting and transcription factor assays. Results AMB cells continued to grow for more than 100 population doublings. Stimulation with TNF ‐ α increased IL ‐6 and MMP ‐9 mRNA expressions, as well as NF ‐ κ B activation. Furthermore, TGF ‐ β and IFN ‐ γ dramatically increased TNF ‐ α ‐mediated expressions of MMP ‐9 and IL ‐6 mRNA , respectively, while those responses were suppressed by NF ‐ κ B inhibitor. Conclusion We established an immortalized AM cell line by hTERT transfection. TNF ‐ α ‐mediated regulation of MMP ‐9 and IL ‐6 via NF ‐ κ B may play an important role in the pathological behaviour of AM s, such as bone resorption.