毛蕊花苷通过上调Max蛋白抑制口腔鳞癌细胞的迁移和侵袭

Verbascoside inhibits migration and invasion of oral squamous cell carcinoma cells by upregulating Max protein

目的:探讨毛蕊花苷对口腔鳞癌细胞迁移和侵袭的影响及其机制。方法:质谱分析空白对照组及毛蕊花苷处理组口腔鳞癌细胞HN4中蛋白的表达谱,筛选毛蕊花苷处理后丰度高且表达显著增加的蛋白,用Western blot验证质谱分析的结果,并在HN4细胞中运用shRNA干扰关键蛋白的表达,分析毛蕊花苷处理对HN4细胞迁移和侵袭的影响。结果:质谱分析结果显示经毛蕊花苷处理后,HN4细胞中Max、TMPRSS11F、ROBO4和AP4E1等18种蛋白表达显著升高(变化倍数>1.5),对其中改变最明显的Max蛋白进行Western blot检测,结果与质谱芯片结果一致;shRNA干扰HN4细胞中Max的表达,qRT-PCR和Western blot检测发现HN4细胞中Max的表达明显下调,运用毛蕊花苷刺激HN4细胞,发现毛蕊花苷可显著抑制HN4细胞的迁移和侵袭。然而,当敲减Max蛋白后,毛蕊花苷抑制HN4细胞迁移和侵袭的能力显著减弱。结论:毛蕊花苷可能通过增强口腔鳞癌细胞中Max的表达抑制口腔鳞癌细胞的迁移和侵袭。

Objective:To analyze the effect and mechanism of verbascoside on the migration and invasion of oral squamous cell carcinoma. Methods:The expression profiles of HN4 protein in oral squamous cell carcinoma cells treated with verbascoside and the blank control group were analyzed by mass spectrometry. Proteins with high abundance and significantly increased expression after verbascoside treatment were screened. The results of mass spectrometry were verified by Western blotting. The expression of key proteins in HN4 cells was interfered by shRNA to analyze verbascoside. Effects of verbascoside on migration and invasion of squamous cell carcinoma HN4 cells were analyzed. Results:The results of mass spectrometry analysis showed that 18 proteins,including Max,MPRSS11 F,ROBO4,AP4 E1,and etc,was significantly increased in oral squamous cell carcinoma HN4 cells after treatment with verbascoside(fold change>1.5). The Max proteins with the most obvious changes were detected by Western blotting. The results of Western blotting were consistent with those of mass spectrometry microarray. shRNA interfered with the expression of Max in oral squamous cell carcinoma HN4 cells. qRT-PCR and Western blotting showed that the expression of Max in HN4 cells was significantly down-regulated. Further stimulation of HN4 cells with verbascoside showed that the migration and invasion of HN4 cells were significantly inhibited. However,the ability of verbascoside to inhibit the migration and invasion of HN4 cells decreased significantly after knockdown of Max protein. Conclusion:It is possible that verbascoside inhibits the migration and invasion of oral squamous cell carcinoma cells by enhancing the expression of Max in oral squamous cell carcinoma cells.