【摘要】 目的 探讨粉防己碱对胶质瘤细胞恶性表型的影响及分子机制。方法 基于TCGA、GEO、MSigDB和CTD数据库筛选胶质瘤中粉防己碱潜在结合的高表达蛋白编码基因，根据单基因GSEA通路分析筛选转化生长因子-β1(TGF-β1)下游通路；行CCK-8实验测定细胞活力，结晶紫染色测定克隆形成能力，流式细胞术分析细胞周期分布及凋亡率，划痕实验和Transwell实验评估细胞迁移侵袭能力；行RT-qPCR和Western blot实验检测上皮-间质转化（EMT）相关蛋白及Smad2-Snail通路调控因子表达；通过转染构建TGF-β1高表达和低表达细胞模型。结果 TGF-β1在胶质瘤患者肿瘤组织中高表达（P<0.05），且同患者不良预后相关，粉防己碱可潜在结合并抑制TGF-β1蛋白；粉防己碱显著抑制胶质瘤细胞U251和T98G细胞的增殖、迁移、侵袭活性，诱导细胞凋亡，并抑制EMT过程，阻断TGF-β1/Smad2-Snail通路激活（P<0.01）；下调TGF-β1表达可抑制胶质瘤细胞增殖，并诱导细胞凋亡（P<0.05,P<0.01）。结论 粉防己碱通过调控TGF-β1/Smad2-Snail轴抑制胶质瘤的EMT过程，为胶质瘤治疗提供了新的理论依据。更多还原

【Abstract】 Objective To investigate the effect of tetrandrine on the malignant phenotype of glioma cells and its molecular mechanism. Methods High-expression protein-coding genes potentially bound by tetrandrine in glioma were screened based on TCGA, GEO, MSigDB and CTD databases. The downstream pathways of transforming growth factor-β 1(TGF-β1) were screened according to single-gene GSEA pathway analysis. CCK-8 assay was performed to determine cell viability, and crystal violet staining was used to measure colony-forming ability. Flow cytometry was applied to analyze cell cycle distribution and apoptosis rate, and wound-healing assay and Transwell assay were carried out to evaluate cell migration and invasion ability. RT-qPCR and Western blot were used to detect the expression of epithelial-mesenchymal transition(EMT)-related proteins and regulatory factors of the Smad2-Snail pathway. Cell models with high and low expression of TGF-β1 were constructed by transfection. Results TGF-β1 was highly expressed in the tumor tissues of glioma patients and was associated with poor prognosis. Tetrandrine could potentially bind to and inhibit TGF-β1 protein(P<0.05). Tetrandrine significantly inhibited the proliferation, migration and invasion activities of glioma cells U251 and T98G, induced apoptosis, inhibited the EMT process and blocked the activation of the TGF-β1/Smad2-Snail pathway(P<0.01). Down-regulation of TGF-β1 expression could inhibit the proliferation of glioma cells and induce apoptosis(P<0.05, P<0.01). Conclusion Tetrandrine inhibits the EMT process of glioma by regulating the TGF-β1/Smad2-Snail axis, providing a new theoretical basis for the treatment of glioma.更多还原