鼠尾草酚抑制巨噬细胞NLRP3/IL-1β 通路活化对口腔鳞癌细胞迁移和侵袭的影响研究

Effect of Carnosol on migration and invasion of human oral squamous carcinoma cells via inhibition of NLRP3/ IL-1β pathway in macrophages

目的旨在探讨巨噬细胞中NLRP3/IL-1β通路对口腔鳞癌细胞迁移和侵袭的影响及鼠尾草酚对这一过程的调控作用。方法qRT-PCR,Western Blot,ELISA法检测口腔鳞癌细胞与巨噬细胞串扰对巨噬细胞NLRP3炎症小体激活及IL-1β释放的影响;Transwell以及qRT-PCR实验检测不同浓度IL-1β对于口腔鳞癌细胞迁移和侵袭能力的影响;ELISA实验检测鼠尾草酚对于串扰引起巨噬细胞NLRP3炎症小体激活及IL-1β释放的影响;Transwell实验检测鼠尾草酚通过影响串扰引起的口腔鳞癌细胞迁移和侵袭能力的改变;CCK8实验检测鼠尾草酚培养口腔鳞癌细胞系Cal-27和单核巨噬细胞系THP-1的毒性作用。结果口腔鳞癌细胞与巨噬细胞串扰导致巨噬细胞中NLRP3炎症小体激活,IL-1β释放;Transwell实验结果表明,IL-1β以剂量依赖性的方式促进口腔鳞癌细胞迁移和侵袭;鼠尾草酚以剂量依赖性的方式抑制着串扰引起的巨噬细胞NLRP3炎症小体激活与IL-1β释放,且40μM浓度的鼠尾草酚显著抑制被串扰增强的口腔鳞癌细胞迁移和侵袭能力;CCK8实验结果表明0~100μM浓度的鼠尾草酚对于Cal-27和THP-1细胞均无毒性。结论本研究证明口腔鳞癌细胞与巨噬细胞的串扰可以促进巨噬细胞中NLRP3炎症小体激活与IL-1β释放,进而显著增强口腔鳞癌细胞的迁移和侵袭能力;鼠尾草酚通过抑制巨噬细胞NLRP3/IL-1β通路的激活与IL-1β释放,抑制口腔鳞癌细胞的迁移和侵袭。

Objective The aim of this study was to investigate the effect of NLRP3/IL-1βpathway in macrophages on the migration and invasion of oral squamous cell carcinoma cells and the regulatory role of Carnosol on this process.Methods qRT-PCR,Western Blot and ELISA were used to detect the effects of Carnosol between oral squamous cell carcinoma cells and macrophages on NLRP3 inflammasome activation and IL-1βrelease in macrophages.Transwell and qRT-PCR assays were used to determine the effects of IL-1βon the migration and invasion of oral squamous cell cells.The effect of carnol on the activation of NLRP3 inflammasome and the release of IL-1βin macrophages induced by crosstalk was detected by ELISA.The transwell assay was used to detect the changes in the migration and invasion ability of oral squamous cell carcinoma cells caused by the effect of the crosstalk.CCK8 assay was used to detect the toxic effects of Carnosol cultured oral squamous cell carcinoma cell line Cal-27 and monocyte macrophage cell line THP-1.Results Crosstalk between oral squamous cell carcinoma cells and macrophages leads to NLRP3 inflammasome activation and IL-1βrelease from macrophages.Transwell assay results suggest that IL-1βpromotes oral squamous cell migration and invasion in a dose-dependent manner.Carnosol inhibits crosstalk-induced NLRP3 inflammasome activation and IL-1βrelease in a dose-dependent manner,and a concentration of 40μM Carnosol effectively inhibits the increased migration and invasion of oral squamous cell carcinoma cells enhanced by the crosstalk;The results of CCK8 experiments show that Carnosol at concentrations of 0-100μM is not toxic to both Cal-27 and THP-1 cells.Conclusion This study demonstrated that the crosstalk between oral squamous cell carcinoma cells and macrophages could promote NLRP3 inflammasome activation and IL-1βrelease from macrophages,thereby significantly enhancing the migration and invasion ability of oral squamous cell carcinoma cells.Carnosol inhibits the migration and invasion of oral squamous cell carcinoma cells by inhibiting the activation of NLRP3/IL-1βpathway and the release of IL-1βin macrophages.