基质金属蛋白酶9抑制剂对人舌鳞癌SCC-25细胞侵袭、迁移能力的影响及机制探讨

Effect of MMP-9 inhibitor on invasion and migration ability of human tongue squamous cell carcinoma SCC-25 cell and its mechanism

目的 :探讨基质金属蛋白酶9(MMP-9)抑制剂对人舌鳞状细胞癌(TSCC)SCC-25细胞侵袭、迁移能力的影响及其相关机制。方法:取对数生长期SCC-25细胞,分别采用0、5、10、20μmol/L的MMP-9抑制剂BB-94干预,分别设为对照组、实验A组、实验B组和实验C组。采用RT-qPCR和Western免疫印迹测定MMP-9 mRNA及蛋白的表达;Transwell小室实验检测细胞侵袭能力;划痕实验测定迁移能力;Western免疫印迹测定细胞间黏附分子-1 (ICAM-1)、E钙黏附蛋白(E-cad)、间隙连接蛋白43(Cx43)的蛋白表达水平。采用SPSS 22.0软件包对数据进行统计学分析。结果:与对照组相比,实验A、B、C组MMP-9 mRNA及蛋白相对表达量、迁移率降低,穿膜细胞数量/视野减少(P<0.05);与实验A组相比,实验B、C组MMP-9 mRNA及蛋白相对表达量、迁移率降低,穿膜细胞数量/视野减少(P<0.05);与实验B组相比,实验C组MMP-9 mRNA及蛋白相对表达量、迁移率降低,穿膜细胞数量/视野减少(P<0.05)。与对照组相比,实验A、B、C组ICAM-1蛋白相对表达量降低,E-cad、Cx43蛋白相对表达量升高(P<0.05);与实验A组相比,实验B、C组ICAM-1蛋白相对表达量降低,E-cad、Cx43蛋白相对表达量升高(P<0.05);与实验B组相比,实验C组ICAM-1蛋白相对表达量降低,E-cad、Cx43蛋白相对表达量升高(P<0.05)。结论:MMP-9抑制剂可抑制TSCC细胞侵袭、迁移,且呈剂量依赖性,推测其作用机制与下调ICAM-1表达,上调E-cad、Cx43表达有关。

PURPOSE: To study the effect of matrix metalloproteinase-9(MMP-9) inhibitors on invasion and migration ability of human tongue squamous cell carcinoma(TSCC) SCC-25 cells, and explore its related mechanisms. METHODS:SCC-25 cells in the logarithmic phase were taken and treated with 0, 5, 10, and 20 μmol/L MMP-9 inhibitor BB-94 respectively, and divided into control group, experimental group A, B and C. RT-qPCR, Western blot were used to determine the expression of MMP-9. Transwell chamber experiment was used to determine invasion ability. Scratch test was used to determine migration ability. Western blot was used to determine the protein expression of intercellular adhesion molecule-1(ICAM-1), E-cadherin(E-cad), and connexin 43(Cx43). SPSS 22.0 software package was used for data analysis. RESULTS: Compared with the control group, the relative expression of MMP-9 mRNA and protein, migration rate,and number of penetrating cells/field of view in experimental A, B, and C group decreased significantly(P<0.05). Compared with experimental group A, the relative expression of MMP-9 mRNA and protein, migration rate, and number of penetrating cells/field of view in experimental B, and C group decreased significantly(P<0.05). Compared with experimental group B, the relative expression of MMP-9 mRNA and protein, migration rate, the number of penetrating cells/field of view in experimental group C decreased(P<0.05). Compared with the control group, the relative expression of ICAM-1 protein in experimental group A, B, and C decreased, while the relative expression of E-cad and Cx43 protein increased significantly(P<0.05). Compared with experimental group A, the relative expression of ICAM-1 protein in experimental group B and C decreased, while the relative expression of E-cad and Cx43 protein increased significantly(P<0.05). Compared with experimental group B, the relative expression of ICAM-1 protein in experimental group C decreased, while the relative expression of E-cad and Cx43 protein increased(P<0.05). CONCLUSIONS: MMP-9 inhibitor can inhibit TSCC cell invasion and migration in a dose-dependent manner. It is speculated that its mechanism of action is related to down-regulation of ICAM-1 expression and up-regulation of E-cad and Cx43 expression.