冬凌草甲素通过根蛋白抑制胰腺癌细胞增殖和迁移的作用机制分析

Mechanistic analysis of oridonin-mediated inhibition of pancreatic cancer cell proliferation and migration via radixin

摘要:

目的 探究冬凌草甲素通过根蛋白（RDX）抑制胰腺癌细胞增殖和迁移的作用机制。方法 采用CCK8法评估冬凌草甲素对人胰腺癌细胞增殖的影响并计算IC50；通过Transwell和划痕试验分析其对细胞迁移能力的影响。结合生物信息学方法和分子实验，分析RDX基因在胰腺癌组织中的表达情况，及其对胰腺癌细胞增殖和迁移的影响。通过基因富集分析和GEPIA数据库分析RDX与下游通路基因的相关性，并通过分子实验进行验证。结果 冬凌草甲素40 μmol/L、80 μmol/L组细胞相对活力较对照组低（P <0.05）。冬凌草甲素组迁移细胞数低于对照组（P <0.05），相对迁移面积小于对照组（P <0.05）。胰腺癌组织RDX基因相对表达量高于正常组织（P <0.05）。冬凌草甲素组RDX基因和蛋白的相对表达量均低于对照组（P <0.05）。si-RDX1组和si-RDX2组RDX基因和蛋白相对表达量均低于si-NC组（P <0.05）。si-RDX1组和si-RDX2组细胞相对活力、迁移细胞数均低于si-NC组（P <0.05），相对迁移面积均小于si-NC组（P <0.05）。基因富集分析结果显示RDX可影响胰腺癌细胞黏附分子的表达（P <0.05），RDX与SNAI1在胰腺癌组织中表达呈正相关性（r =0.550，P <0.05）。冬凌草甲素组SNAI1基因和蛋白相对表达量均低于对照组（P <0.05）。si-RDX1组和si-RDX2组SNAI1基因和蛋白相对表达量均低于si-NC组（P <0.05）。结论 冬凌草甲素可能通过降低RDX基因表达，抑制胰腺癌PANC-1细胞增殖和迁移。

Abstract:

Objective To explore the mechanism by which oridonin inhibits the proliferation and migration of pancreatic cancer cells through radixin (RDX).Methods The CCK8 assay was used to detect the effects of oridonin on the proliferation of PANC-1 pancreatic cancer cells, and the IC50 was calculated. The effect of oridonin on cell migration was assessed using Transwell and wound healing assays. A combination of bioinformatics analysis and molecular experiments was used to investigate the expression of the RDX gene in pancreatic cancer tissues and its effect on the proliferation and migration of pancreatic cancer cells. Single gene enrichment analysis and the GEPIA database were used to analyze the correlation between RDX and downstream pathway genes, and the results were verified by molecular experiments.Results Cells treated with oridonin at 40 μM and 80 μM showed significantly reduced relative viability compared with the control group (P < 0.05). The number of migrating cells and the relative migration area were also lower in the oridonin-treated groups than in controls (P < 0.05). RDX gene expression was significantly higher in pancreatic cancer tissues compared with normal tissues (P < 0.05). Treatment with oridonin significantly reduced both the relative mRNA and protein expression levels of RDX compared with the control group (P < 0.05). In cells transfected with si-RDX1 and si-RDX2, RDX mRNA and protein levels were significantly lower than in the si-NC group (P < 0.05). These knockdown cells also exhibited reduced relative viability, fewer migrating cells, and smaller relative migration areas compared with si-NC-transfected cells (P < 0.05). GSEA results indicated that RDX may regulate the expression of adhesion molecules in pancreatic cancer cells (P < 0.05). Moreover, RDX expression was positively correlated with SNAI1 expression in pancreatic cancer tissues (r = 0.550, P < 0.05). Both mRNA and protein levels of SNAI1 were significantly lower in the oridonin-treated group compared with controls (P < 0.05), and in si-RDX1 and si-RDX2 groups compared with the si-NC group (P < 0.05).Conclusion Oridonin may inhibit the proliferation and migration of PANC-1 pancreatic cancer cells by downregulating RDX gene expression.