大黄素对黑素瘤细胞B16F10迁移的影响

Effects of Emodinon on Migration of B16F10 Melanoma Cells

目的研究大黄素对黑素瘤细胞B16F10的迁移能力及NLRP3炎症小体相关蛋白表达的影响。方法 CCK-8试验、划痕愈合试验及Transwell迁移试验检测大黄素对黑素瘤细胞B16F10活性和迁移能力的影响,Western blot法检测大黄素对黑素瘤细胞B16F10中NLRP3炎症小体相关蛋白表达的影响。结果 CCK-8试验显示,与对照组相比,使用不同浓度大黄素处理24 h及48 h后,黑素瘤B16F10细胞的细胞活性受到明显抑制(P<0.05)。划痕愈合试验结果显示,与对照组相比,使用20、40、60μmol/L大黄素处理后,黑素瘤B16F10细胞的划痕愈合能力显著下降。Transwell试验显示,与对照组相比,使用20、40、60μmol/L大黄素处理24 h后,穿过隔膜的细胞明显减少(P<0.05),提示大黄素可以抑制黑素瘤B16F10细胞的迁移能力。Western blot结果显示,与对照组相比,使用20、40、60μmol/L大黄素处理24 h后,NLRP3蛋白的表达水平明显下降。结论大黄素可抑制黑素瘤细胞B16F10细胞的迁移能力,其机制可能与NLRP3炎症小体相关蛋白的下调有关。

Objective To investigate the effects of emodin on the migration of human B16 F10 melanoma cells and the expression of NLRP3.Methods CCK-8 assay,wound healing assay and transwell assay were used to detect the effect of emodin on the cell viability and migration ability of B16 F10 cells.Western blot was used to detect the effect of emodin on the expression of NLRP3 in B16 F10 melanoma cells.Results CCK-8 assay showed that emodin reduced cell viability(P<0.05)compared with the control group after treatment with various concentrations of emodin for 24 hours and 48 hours.Results of wound healing assays showed that the wound healing area was significantly reduced compared with the control group after treatment with 20,40 and 60 μmol/L of emodin.Transwell assay showed that the number of transmembrane cells was decreased significantly after being treated with 20,40 and 60 μmol/L of emodin for 24 hours(P < 0.05),indicating that emodin was able to inhibit cell migration of B16 F10 cells.The result of Western blot showed that the expression level of NLRP3 protein was decreased significantly after treatment with 20,40 and 60 μmol/L of emodin.Conclusion Emodin can inhibit the migration ability of B16 F10 cells,which may involve the down-regulation of inflammasome relative proteins including NLRP3.