Objective: To investigate the inhibitory effect of Doxycycline on the adhesion of melanoma cells and its mechanism.Methods: Melanoma B16F110, WM351 and WM451 cell lines were treated with Doxycycline at different doses and through different administration methods.MMT, cell adhesion experiment and Western blot were used to observe the effect of different treatment.A total of 20 C57/BL mice were transplanted with melanoma through injection of B16F10 cells and were divided into two groups.One group was treated with Doxycycline, and the other NaCl solution.Carcinoma tissues were obtained and underwentWestern blot and gelatin zymography.Results: MTT analysis showed that the inhibition ratio was significantly higher in the group receiving treatment before cell adhesion than in the group receiving treatment after cell adhesion ( P ＜ 0.05 ).Cell adhesion experiment showed that Doxycycline had an inhibitory effect on the adhesion of B16F110, WM351 and WM451 cells, with a statistical significance ( P ＜ 0.05 ).Western blot revealed that FAK expression in the three cell lines was decreased at 12 h after Doxycycline treatment.Observations from the mice melanoma model suggested obvious changes in FAK expression and its phosphorylation level ( P ＜ 0.05 ).Conclusion: Doxycycline can interfere the adhesion of melanoma cells, possibly through inhibiting FAK expression and its phosphorylation.%目的:研究多西环素对黑色素瘤细胞黏附的抑制作用及分子机制.方法:对黑色素瘤细胞系B16F10、WM351、WM451分别以不同给药方式和给药浓度进行处理,利用MTT实验、细胞黏附实验和Western blot技术检测比较其差异.C57/BL小鼠20只进行黑色素瘤动物实验,接种B16F10后随机分为2组,分别给予多西环素和NaCl溶液处理,取瘤组织进行Western blot和明胶酶谱检测.结果:MTT实验显示"贴壁前"给药组的细胞抑制率显著高于"贴壁后"给药组且差异具有统计学意义(P<0.05).细胞黏附实验分析显示多西环素对3种黑色素瘤细胞均有抑制黏附作用,差异有统计学意义(P<0.05).Western blot结果显示利用多西环素处理12h后,3种细胞均出现了不同程度的FAK表达水平降低.荷瘤小鼠模型结果表明FAK及其磷酸化水平均有明显改变(P<0.05).结论:多西环素可能通过抑制黑色素瘤细胞FAK及其磷酸化,从而干扰肿瘤细胞的黏附作用,进而引发一系列的后续药理作用.
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