二甲双胍对口腔鳞癌细胞迁移的体外作用研究

Effect of metformin on migration of oral squamous cell carcinoma cells in vitro

目的:从体外细胞水平研究二甲双胍对口腔鳞癌细胞迁移能力的影响,并初步探讨可能的机制和潜在价值。方法:复苏培养口腔鳞癌细胞系SCC4和CAL27,选用含不同浓度(0、2、5、10、20 mmol/L)二甲双胍培养基处理培养SCC4和CAL27细胞,CCK⁃8法测定细胞活性。选定二甲双胍低细胞毒性浓度后采用细胞平板克隆、细胞划痕实验测定细胞增殖和迁移能力,明胶酶谱法测定口腔鳞癌细胞的细胞基质金属蛋白酶⁃2/9(matrix metalloprotein⁃ase⁃2/9,MMP⁃2/9)的外分泌能力,免疫印迹法对口腔鳞癌细胞内与上皮⁃间充质转化(epithelial mesenchymal transi⁃tion,EMT)相关的蛋白表达情况进行测定。结果:低细胞毒性浓度的二甲双胍对口腔鳞癌细胞体外增殖和迁移有显著抑制作用(P<0.05)。二甲双胍能够显著抑制口腔鳞癌细胞的外分泌MMP⁃2/9的能力(P<0.05)。二甲双胍能显著调控口腔癌细胞中EMT相关通路标志蛋白的表达(P<0.05),抑制其信号转导。结论:二甲双胍可通过下调MMP⁃2/9外分泌和调控EMT相关信号转导抑制口腔癌细胞迁移。

Objective:To study the effect of metformin on the migration ability of oral squamous cell carcinoma cells in vitro,and to explore the possible mechanism and potential value.Methods:The oral squamous cell carcinoma cell lines SCC4 and CAL27 were resuscitated and cultured.Firstly,cells were treated with different concentrations of metformin(0,2,5,10,20 mmol/L)and then determined low cytotoxic concentration via cell viability assay by CCK⁃8.Then,the ability of cell proliferation and migration was determined by cell plate cloning experiment and cell scratch experiment,the exocrine ability of matrix metalloproteinase⁃2/9(MMP⁃2/9)in oral squamous cell carcinoma cells was determined by gelatin zymography,and expression of proteins related to epithelial⁃mesenchymal transition(EMT)in oral squamous cell carcinoma cells was detected by Western Blotting.Results:Metformin at low cytotoxic concentration could significantly inhibit the proliferation and migra⁃tion of oral squamous cell carcinoma cells in vitro(P<0.05).Metformin could significantly inhibit the expression of exocrine MMP⁃2/9 in oral squamous cell carcinoma cells.Metformin can inhibit EMT signaling pathway via significantly regulating re⁃lated marker proteins(P<0.05).Conclusion:Metformin can inhibit the migration of oral cancer cells by down⁃regulating MMP⁃2/9 exocrine and regulating EMT⁃related signal transduction.