白头翁汤通过降低BUB1表达抑制食管癌细胞生长的研究

Baitouweng decoction suppresses the growth of esophageal carcinoma cells by reducing BUB1 expression

目的探讨白头翁汤通过降低BUB1表达抑制食管癌细胞增殖的作用机制。方法运用基因芯片技术筛选出食管癌组织和正常组织差异表达基因,并用生物信息学方法分析差异表达基因。将EC9706细胞分为空白对照组、白头翁汤组、si-BUB1组和白头翁汤+si-BUB1组。通过MTT法检测细胞增殖活力;用流式细胞术测定细胞周期和凋亡;通过qRT-PCR测定BUB1 mRNA的表达水平;通过蛋白质印迹测量BUB1、STAT3、p-STAT3、CCNB1、CDK1、Bcl-2、Caspase-3和Caspase-9蛋白水平;用划痕和Transwell小室实验测定细胞的迁移和侵袭能力。结果差异表达基因参与生物学过程、信号通路和网络构建主要与细胞有丝分裂相关,BUB1是关键的核心基因,白头翁汤抑制BUB1表达。此外,体外实验表明,白头翁汤显著抑制食管癌细胞的生长、迁移和侵袭,诱导细胞凋亡和G2/M期阻滞。进一步分析表明,该方剂可能通过下调BUB1表达水平来抑制食管癌细胞的生长。白头翁汤处理后,食管癌细胞中Caspase-和Caspase-9表达量升高,而BUB1、CCNB1、CDK1、p-STAT3和Bcl-2表达量显着降低。STAT3信号通路也被发现在此过程中发挥着重要作用。结论白头翁汤可能通过降低BUB1表达抑制食管癌细胞的生长。

Objective To investigate the mechanism of Baitouweng decoction inhibiting the proliferation of esophageal carcinoma cells by decreasing the expression of BUB1.Methods The differentially expressed genes in EC tissues and normal tissues were screened by the cDNA microarray technique.The differentially expressed genes were analyzed by bioinformatics methods.We divided EC9706 cells into blank control group,Baitouweng decoction,si-BUB1 group and Baitouweng decoction+si-BUB1 group.We detected the activity of EC9706 cells by the MTT method.Cell cycle and apoptosis were measured by flow cytometry.The expression of BUB1 was measured by qRT-PCR.The levels of BUB1,STAT3,p-STAT3,CDK1,Bcl-2,Caspase3 and Caspase9 were measured by Western blotting.The migration and invasion abilities of the cells were measured by wound-healing and Transwell invasion assays.Results Differentially ex⁃pressed genes are involved in biological processes,signaling pathways and network construction,which are mainly related to mitosis.Bub1 is the key hub gene,and Baitouweng decoction inhibits BUB1 expression.Moreover,in vitro experiments demonstrated that Baitou⁃weng decoction significantly inhibited the growth,migration,and invasiveness of esophageal carcinoma cells and induced cell apoptosis and G2/M phase arrest.Further analysis showed that the decoction may inhibit the growth of esophageal carcinoma cells by downregulat⁃ing the BUB1 expression level.After Baitouweng decoction treatment,the expression levels of Caspase-3 and Caspase-9 in esophageal carcinoma cells were increased,while the expression of BUB1,CCNB1,CDK1,p-STAT and Bcl-2 was significantly decreased.The STAT3 signaling pathway was also found to play an important role in this process.Conclusion Baitouweng decoction may inhibit the growth of esophageal carcinoma cells by decreasing the expression of BUB1.