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Wnt5A exerts immunomodulatory activity in the human ovarian cancer cell line SKOV‐3
Author(s) -
Arabzadeh Somayeh,
Hossein Ghamartaj,
Zarnani Amir Hassan
Publication year - 2016
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1002/cbin.10551
Subject(s) - cytokine , transfection , chemotaxis , chemokine , cell culture , cell , small interfering rna , tumor necrosis factor alpha , chemistry , microbiology and biotechnology , inflammation , biology , cancer research , immunology , biochemistry , receptor , genetics
The purpose of this study was three fold: (1) to reveal the implications of Wnt5A for cytokine and chemokine production by human ovarian cancer cell line SKOV‐3 cells, (2) to determine the influence of Wnt5A on chemotactic SKOV‐3 cell migration, and (3) to assess the effect of inflammatory mediators on Wnt5A expression levels and to describe its underlying molecular mechanisms. A cytokine array was performed using a conditioned medium harvested from SKOV‐3 cells transfected with specific siRNAs against Wnt5A or with scrambled siRNA and a transfection reagent alone as negative controls for 48 h. Chemotactic cell migration was performed using transwells. Inflammation‐induced Wnt5A expression was determined by treating cells with recombinant human (rh) IL‐1β, IFNβ, or TNFα alone or in combination with STAT3 and NF‐κB inhibitors for different time durations. The cytokine array showed the suppression of GCSF, GM‐CSF, IL‐1α, IL‐2, IL‐13, and MCP‐3 production, whereas cell RANTES and IL‐7 showed increased levels in Wnt5A knock‐down cells compared with those in controls. Chemotactic migration decreased significantly when the conditioned medium from Wnt5A knock‐down cells was applied to the upper chamber of the transwell. Compared with the control, there were 30‐fold and five‐fold increases in Wnt5A mRNA levels in cells treated, with rhIL‐1β and rhIFNβ, respectively after 8 h ( P < 0.001). Compared with the control, TNF‐α had a 1.8‐fold increased levels of Wnt5A mRNA after 4 h ( P < 0.01). Both NF‐κB and STAT3 inhibitors decreased inflammation‐induced Wnt5A expression. This study revealed a previously unrecognized immunomodulatory role of endogenous Wnt5A in ovarian cancer cells, which could further influence chemotactic cell migration.