毛兰素通过TNF-α信号通路抑制食管癌细胞增殖

Erianin inhibits the proliferation of esophageal cancer cells through TNF-α signaling pathway

为探究毛兰素介导TNF-α信号通路对食管癌细胞增殖的影响,应用网络药理学分析毛兰素抗食管癌作用靶点,以不同浓度毛兰素(0、15、30、45和60 nmol/L)处理食管癌EC109、KYSE510细胞,采用CCK-8试验、克隆形成试验检测细胞增殖能力;构建食管癌裸鼠模型,检测瘤体组织Ki67蛋白表达;采用Western blotting检测TNF-α及其下游通路相关蛋白表达;应用生物信息学方法分析TNF-α在食管癌中的表达情况及其对细胞增殖的影响。结果显示,TNF-α是毛兰素抗食管癌作用靶点中的Top1基因;与0 nmol/L毛兰素比较,经15、30、45和60 nmol/L毛兰素处理后,EC109、KYSE510细胞增殖率,克隆形成率,磷酸化(phosphorylated,p)-NF-κB p65、p-MAPK、p-JNK及TNF-α蛋白表达水平显著下降(P<0.05);与对照组比较,毛兰素组小鼠瘤体体积、质量及瘤体组织中Ki67蛋白表达显著下降(P<0.05);TNF-α在食管癌高表达,与细胞增殖和周期相关的基因集均明显富集在TNF-α高表达组,与细胞凋亡相关的基因集均明显富集在TNF-α低表达组;加入TNF-α后,EC109、KYSE510细胞的活力、增殖能力显著上升(P<0.05),毛兰素抑制NF-κB信号通路激活的现象明显被逆转。该研究提示,毛兰素可抑制食管癌细胞增殖,抑制移植瘤生长,其机制可能与抑制TNF-α及其下游信号通路有关。

In order to explore the effects of erianin on the proliferation of esophageal cancer cells through TNF-αsignaling pathway,network pharmacological analysis was used to identify the target of erianin against esophageal cancer.Esophageal cancer cells EC109 and KYSE510 were treated with different concentrations of erianin(0,15,30,45,60 nmol/L),and cell proliferation was detected by CCK-8 and clone formation assays.Nude mice were used to construct the esophageal cancer model.The expression of Ki67 protein in tumor tissue was detected and expressions of TNF-αand its downstream pathway related proteins were examined by Western blotting.The expression of TNF-αin esophageal cancer and its effects on cell proliferation were analyzed by bioinformatics.The results showed that TNF-αwas the Top1 gene in the anti-esophageal cancer target of erianin.Compared to those of the 0 nmol/L erianin group,the cell proliferation rates of EC109 and KYSE510,clone formation rate,and expression levels of phosphorylated(p)-NF-κB p65,p-MAPK,p-JNK,and TNF-αprotein were significantly decreased after treatment with erianin(15,30,45,60 nmol/L,P<0.05).Compared to those of the control group,the volume and mass of tumors,and expression of Ki67 protein in the erianin-treated tumor tissue were significantly decreased(P<0.05).The expression of TNF-αwas up-regulated in esophageal cancer.The gene sets related to cell proliferation and cycle were significantly enriched in TNF-αhigh-expression group,while the gene sets related to apoptosis were significantly enriched in TNF-αlow-expression group.After treatment with TNF-α,cell viability and proliferation of EC109 and KYSE510 were significantly increased(P<0.05)and the phenomenon that erianin inhibited the activation of NF-κB signaling pathway was significantly reversed.This study concludes that erianin can inhibit the proliferation of esophageal cancer cells and the growth of transplanted tumors by inhibiting the TNF-αand its downstream signaling pathway.