FUT4-siRNA增强5-aza-dC对鳞癌细胞增殖和迁移的抑制

FUT4- siRNA Increased 5-aza-d C Induced Inhibition on Proliferation and Migration in Squamous Carcinoma Cells

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 for the synthesis of fucosylated oligosaccharides. RNAi of FUT4 has been reported to suppress the proliferation of squamous carcinoma cells. 5-aza-d C is a common drug for squamous carcinoma chemotherapy. However,it is unknown whether FUT4-siRNA could be used in combination with 5-aza-d C treatment. In this study,the effect of5-aza-d C combined FUT4-siRNA treatment on squamous carcinoma proliferation and migration were investigated in A431 and SCC12 cells. The results of MTT and flowcytometry assays showed that 18%and 20% of growth inhibition were observed in A431 and SCC12 cells treated with 5 μmol / L 5-aza-d C for4 days（ P 0. 05）,respectively. The cell percentage were decreased in G1 phase and increased in S phase. Western blotting showed that the FUT4 expression was lower,but more significantly increased with 5-aza-d C treatment,in SCC12 than in A431 cells. The results from trypan blue staining and woundhealing assays indicated that FUT4-siRNA transfection decreased FUT4 expression and amplified the 5-aza-d C inhibition of cell proliferation（ 54% and 60% in A431 and SCC12） and migration. These suggested that FUT4-siRNA increased the proliferation inhibition by 5-aza-d C treatments in squamous carcinoma cells,which involved in cell cycle arrest at S phase.