莱菔硫烷对人肾癌细胞增殖、迁移和侵袭的影响及作用机制

The effects of sulforaphane on proliferation,migration and invasion of human renal carcinoma cells and its mechanism

目的探讨莱菔硫烷(SFN)对人肾癌细胞的增殖、迁移和侵袭的影响及作用机制。方法体外培养人肾癌细胞786-O,并将其分为4组:空白对照组、SFN低浓度组(5μmol/L)、SFN中浓度组(10μmol/L)、SFN高浓度组(20μmol/L),用CCK-8检测肾癌细胞786-O增殖活化的情况;分别用细胞划痕、Transwell迁移实验观察SFN对肾癌细胞786-O迁移能力的影响;用Transwell侵袭实验观察SFN对肾癌细胞786-O侵袭能力的影响;运用Western blot和qRT-PCR方法检测SFN对上皮间质转化(EMT)相关蛋白和mRNA表达的影响;用Western blot检测SFN对NF-κB信号通路相关蛋白表达的影响。结果SFN处理24、48、72 h后,肾癌细胞786-O的增殖活性随着SFN浓度的升高而下降;与对照组相比,SFN处理组的肾癌细胞迁移和侵袭能力均显著降低;此外,随着SFN浓度的升高,肾癌细胞786-O中E-cadherin的mRNA及蛋白表达水平逐渐升高,而N-cadherin、Vimentin的mRNA及蛋白表达水平随之降低;NF-κB信号通路相关蛋白磷酸化的p65和磷酸化的IκBα的蛋白水平随SFN浓度增加而降低。结论SFN能够显著抑制人肾癌细胞的增殖、迁移和侵袭的能力,其可能通过抑制NF-κB信号通路进而阻碍肾癌的EMT进程来发挥抗肿瘤作用。

Objective To investigate the effects of sulforaphane(SFN)on proliferation,migration and invasion of human renal carcinoma cells and its mechanism.Methods The cultured human renal carcinoma cells 786-O were divided into control group(0μmol/L)and SFN group(5,10,20μmol/L).The activated proliferation of cells was detected by CCK-8;the effect of SFN on migration of 786-O cells was detected by scratch healing assay and Transwell cell migration assay;the effect of SFN on the invasion ability of 786-O cells was detected by Transwell cell invasion ability assay;Western blot and qRT-PCR were used to detect the effects of SFN on the expression of epithelial-mesenchymal transition(EMT)-related proteins and mRNA.The effect of SFN on the expression of NF-κB signaling pathway was detected by Western blot.Results After SFN treatment for 24,48 and 72 h,the proliferation activity of 786-O cells decreased with the increase of SFN concentration;compared with the control group,the cell migration and invasion ability of SFN-treated group were significantly reduced;with the increase of SFN concentration,the mRNA and protein expression levels of E-cadherin in 786-O cells increased,while the mRNA and protein expression levels of N-cadherin and Vimentin decreased;the levels of NF-κB signaling pathway related protein phosphorylated p65 and phosphorylated IκBαdecreased with the increase of SFN concentration.Conclusion SFN may inhibit the proliferation,migration and invasion of human renal carcinoma cells by regulating the EMT process of renal carcinoma through inhibition of NF-κB signaling pathway.