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岩藻糖基转移酶IV(fucosyltransferase IV,FUT4)是催化蛋白质岩藻糖基化的关键酶.已经证明,FUT4-siRNA能够抑制鳞癌细胞的增殖.5-氮杂-2-脱氧胞苷(5-aza-d C)是临床常用化疗药物,但5-aza-d C是否对鳞癌有治疗作用,以及与FUT4-siRNA联合使用能否加强对鳞癌细胞增殖和迁移的抑制尚不清楚.本研究以鳞癌细胞系A431和SCC12为对象,探讨5-aza-d C及其与FUT4-siRNA联合使用对细胞增殖和迁移的影响.MTT结合流式细胞周期分析显示,5-aza-d C处理A431和SCC12细胞4 d后,细胞增殖被明显抑制,抑制率分别为18%和20%(P<0.05);与对照组比较,加药处理组G1期细胞数量减少,S期细胞数量明显增加.Western印迹结果揭示,A431细胞FUT4表达水平较SCC12细胞高.经5-aza-d C处理后SCC12细胞FUT4表达有所增加,但仍低于A431细胞中的表达.FUT4-siRNA转染结合台盼蓝活细胞记数证明,FUT4-siRNA明显降低细胞FUT4表达,5-azad C处理同时转染FUT4-siRNA的A431和SCC12细胞增殖进一步被抑制,抑制率分别为54%和60%(P<0.05).细胞划痕法显示,5-aza-d C与FUT4-siRNA联合使用,对细胞迁移能力的抑制作用比5-aza-d C单独使用增强.上述结果提示,5-aza-d C通过诱导细胞S期阻滞抑制鳞癌细胞增殖,FUT4-siRNA与5-aza-d C联合使用可加强对细胞增殖和迁移的抑制.
Fucosyltransferase IV (FUT4) is a key enzyme that catalyzes the fucosylation of proteins.FUT4-siRNA has been shown to inhibit the proliferation of squamous cell carcinoma.5-aza-2-deoxycytidine 5-aza-d C) is a commonly used chemotherapeutic drug in clinic. However, whether 5-aza-d C has a therapeutic effect on squamous cell carcinoma and whether its combination with FUT4-siRNA enhances inhibition of proliferation and migration of squamous cell carcinoma is unclear In this study, we investigated the effects of 5-aza-d C and its combination with FUT4-siRNA on the proliferation and migration of squamous cell carcinoma cell lines A431 and SCC12.MTT combined with flow cytometry analysis showed that 5-aza- Compared with the control group, the number of cells in G1 phase decreased and the number of S phase cells decreased significantly in dC-treated A431 and SCC12 cells for 4 days (P <0.05) The number of FUT4 in A431 cells was significantly higher than that in SCC12 cells.FUT4 expression in SCC12 cells was increased but still lower than that in A431 cells after 5-aza-d C treatment.FUT4-siRNA transfection Staining combined with trypan blue cells showed that FUT4-siRNA significantly reduced the expression of FUT4 in cells, and 5-azad C transfection of FUT4-siR NA of A431 and SCC12 cell proliferation was further inhibited, the inhibition rates were 54% and 60% (P <0.05) .Cell scratch assay showed that 5-aza-d C and FUT4-siRNA combined use of cell migration ability The inhibitory effect was stronger than that of 5-aza-d C. The above results suggest that 5-aza-d C can inhibit the proliferation of squamous cell carcinoma by inducing S phase arrest, and the combination of FUT4-siRNA and 5-aza-d C can be enhanced Inhibition of cell proliferation and migration.