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Upregulation of IL ‐6 expression in human salivary gland cell line by IL ‐17 via activation of p38 MAPK , ERK , PI 3K/Akt, and NF ‐κB pathways
Author(s) -
Wei Lili,
Xiong Haofei,
Li Wei,
Li Bo,
Cheng Yong
Publication year - 2018
Publication title -
journal of oral pathology and medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.887
H-Index - 83
eISSN - 1600-0714
pISSN - 0904-2512
DOI - 10.1111/jop.12765
Subject(s) - mapk/erk pathway , downregulation and upregulation , p38 mitogen activated protein kinases , microbiology and biotechnology , protein kinase b , western blot , kinase , cell culture , biology , signal transduction , chemistry , biochemistry , genetics , gene
Background The human salivary gland ( HSG ) cell line has so far been used as in vitro models for study of the influence of cytokines and pharmacologic agents on salivary glands, as well as a model system for inflammation in Sjögren's syndrome ( SS ). This study aimed to determine the effect of IL ‐17 on IL ‐6 production and the underlying molecular mechanism regulated by the HSG cell line. Methods Immunofluorescence analyses, RT ‐ PCR , and Western blot were conducted to evaluate the IL ‐17 receptor ( IL ‐17R) expression in cultured HSG cells. Real‐time PCR and ELISA were then utilized to establish the mRNA and protein levels of IL ‐6 in IL ‐17‐stimulated HSG cells. Western blot, flow cytometry, immunofluorescence, and inhibitor analyses were conducted to elucidate the involved signaling pathways. Results The HSG cells reliably expressed the IL ‐17R mRNA and its encoded surface‐bound protein. The expression of IL ‐6 mRNA and protein was upregulated by stimulation of HSG cells with IL ‐17; this effect was impeded by IL ‐17‐ or IL ‐17R‐neutralizing antibodies. IL ‐17 stimulation ended up with the fast phosphorylation of p38 mitogen‐activated protein kinase ( MAPK ), extracellular signal‐regulated kinase ( ERK ), Akt, and translocation of nuclear factor‐kappaB ( NF ‐κB) in the HSG cells. p38 MAPK , Akt, and NF ‐κB inhibitors significantly subdued IL ‐6 generation in HSG cells stimulated by IL ‐17. PD 98059, an ERK inhibitor, decreased IL ‐6 generation under low dose of IL ‐17 but not with high dose. Conclusions The HSG cells expressed IL ‐17R and reacted to IL ‐17 to generate IL ‐6 via the stimulation of ERK , p38 MAPK , Akt, and NF ‐κB signaling pathways.