6-姜烯酚通过干预中性粒细胞胞外诱捕网抑制乳腺癌侵袭作用研究OA

Objective To investigate the effect of tumor-associated neutrophils(TANs)in breast cancer 4T1 cells,and the intervention role of the dried ginger active ingredient 6-shogaol on TANs.Methods Mouse neutrophils were isolated and identified,an in vitro co-culture system of neutrophils and tumor cells was established.Effects of TANs and 6-shogaol-treated TANs on 4T1 cell viability,adhe-sion,migration,and invasion were detected via MTT,adhesion,wound healing,and transwell assays.MPO and Citrullinated Histone H3(Cit-H3)was detected to assess NETs release in neutrophils stimulated with PMA or tumor cells,and the effect of tumor cells treated with 6-shogaol on the release of NETs was observed by laser confocal microscopy.ELISA was used to detect TNF-α,IL-6,and IL-1β levels in 6-shogaol-treated TANs.Effect of 6-shogaol on NETs trapping tumor cells was detected by Transwell chamber assay.Results Mouse neutrophils were successfully isolated and purified.TANs promoted 4T1 cell viability,adhesion,migration,and inva-sion,while 6-shogaol reversed these promoting effects by modulating TANs.Results of ELISA assay showed that 6-shogaol reduced the secretion of TNF-α,IL-6 and IL-1β in TANs.The expression of MPO and Cit-H3 in neutrophils was stimulated by PMA or tumor cells,and 6-shogaol inhibited tumor cell-induced excessive NETs release in neutrophils.Transwell assay showed 6-shogaol reduced tumor cell trapping by PMA-induced neutrophil NETs.Conclusion TANs promote the abilities of proliferation,adhesion,migration and invasion of 4T1 cells;6-shogaol reverses promoting effects via intervening in neutrophil recruitment and excessive release of NETs,ameliorates the tumor inflammatory microenvironment and inhibits breast cancer cell invasion and migration.