乳酸对舌鳞癌细胞迁移和侵袭的影响及其机制初探

The effect of lactic acid on the migration and invasion ability of tongue squamous cell carcinoma cells and its preliminary mechanism investigation

目的探究乳酸对人舌鳞癌细胞SCC15迁移和侵袭能力的影响并初步探索机制。方法使用10、15、20 mmol/L乳酸处理舌鳞癌细胞SCC15,采用划痕实验和Transwell迁移实验检测SCC15迁移能力;采用Matrigel侵袭实验检测SCC15侵袭能力;使用10、15 mmol/L乳酸处理舌鳞癌细胞SCC15,采用细胞-细胞黏附实验、细胞-基底黏附实验检测细胞黏附能力;采用免疫荧光、免疫印迹检测磷酸化黏着斑激酶(phosphorylated focal adhesion kinase,p-FAK)表达情况;采用实时荧光定量PCR(Real-time PCR,qRTPCR)检测上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关基因表达情况。结果10 mmol/L和15 mmol/L乳酸可增强SCC15细胞的迁移及侵袭能力(P<0.01),细胞黏附功能如细胞-细胞黏附、细胞-基底黏附水平(P<0.01)及p-FAK表达(P<0.01)均显著下降,EMT相关基因表达显著上调(P<0.05)。结论初步研究发现乳酸能够上调SCC15 EMT相关基因表达,降低细胞黏附水平,提高细胞迁移、侵袭能力。

Objective To investigate the effect of lactic acid on the migration and invasion ability of human tongue squamous cell carcinoma cells SCC15 and explore the underlying preliminary mechanism.Methods After treating SCC15 with lactic acid at 10,15,20 mmol/L,scratch healing test and Transwell assay was utilized to detect the migration ability of SCC15.Matrigel invasion assay was employed to detect the invasion ability of SCC15.After treating SCC15 with lactic acid at 10,15 mmol/L,cell adhesion ability was evaluated through cell-cell adhesion assay and cell-substrate adhesion assay.The expression of phosphorylated focal adhesion kinase(p-FAK)was examined using immunofluorescence and western-blotting.Expression of epithelial-mesenchymal transition(EMT)related genes were measured using real-time quantitative PCR(qRT-PCR).Results Lactic acid enhanced the migration and invasion ability of SCC15(P<0.01),reduced cell-cell adhesion and cell-substrate adhesion levels(P<0.01),downregulated the expression of p-FAK(P<0.01),and upregulated the expression of EMT-related genes(P<0.05).Conclusion Preliminary findings suggest that lactic acid may enhance the migration and invasion ability of SCC15 by upregulating the expression of EMT-related genes.