【摘要】 目的 研究去氢粗毛甘草素(dehydroglyasperin, DHG)对Hep-2细胞(一种喉癌模型细胞)增殖、凋亡、细胞周期及自噬等恶性表型的抑制作用，并探讨该过程中涉及的潜在机制。方法 以人喉鳞状细胞癌Hep-2细胞为模型，通过细胞增殖实验、凋亡检测及细胞周期分析评估DHG的抗肿瘤活性；利用免疫细胞化学(immunocytochemistry, ICC)和Western blot检测自噬标志物微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3B,LC3B)、自噬关键调控蛋白Beclin-1及自噬底物P62蛋白表达。结合qRT-PCR和Western blot分析血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)/磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase, PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路关键分子表达及磷酸化水平变化。结果 DHG显著抑制Hep-2细胞增殖，并呈现剂量和时间依赖性。细胞周期检测显示，DHG诱导S期阻滞，同时促进细胞凋亡。自噬相关标志物LC3B-Ⅱ和Beclin-1表达显著上调，而P62蛋白水平明显降低，提示DHG激活细胞自噬。此外，DHG通过抑制VEGFA/PI3K/AKT信号通路，降低VEGFA表达及VEGFR2、PI3K、AKT的磷酸化水平，进而阻断下游促生存信号传导。结论 去氢粗毛甘草素可以诱导喉癌细胞自噬与凋亡，并阻滞细胞周期进程，其抗肿瘤效应与VEGFA/PI3K/AKT通路活性下调显著相关。更多还原

【Abstract】 OBJECTIVE To investigate the inhibitory effect of dehydroglyasperin(DHG) on the malignant phenotype of laryngeal cancer cells and explore the potential mechanisms involved in this process. METHODS Human laryngeal squamous cell carcinoma Hep-2 cells were used as a model to evaluate the antitumor activity of DHG by cell proliferation assay, apoptosis assay and cell cycle analysis. Immunocytochemistry(ICC) and Western blot were used to detect the expression of autophagy marker microtubule-associated protein 1 light chain 3B(LC3B), key autophagy regulatory protein Beclin-1 and autophagy substrate P62 protein. qRT-PCR and Western blot were used to analyze the expression and phosphorylation levels of key molecules in the vascular endothelial growth factor A(VEGFA)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway. RESULTS DHG significantly inhibited the proliferation of Hep-2 cells in a dose-and time-dependent manner. Cell cycle detection showed that DHG induced S phase arrest and promoted cell apoptosis. The expression of autophagy-related markers LC3B-Ⅱ and Beclin-1 was significantly upregulated, while the level of P62 protein was significantly decreased, indicating that DHG activated cell autophagy. In addition, DHG inhibited the VEGFA/PI3K/AKT signaling pathway, reduced the phosphorylation levels of VEGFR2, PI3K and AKT, and thus blocked the downstream pro-survival signal transduction. CONCLUSION Dehydroglyasperin can induce autophagy and apoptosis and arrest cell cycle progression in laryngeal cancer cells. The anti-tumor effect of dehydroglyasperin is significantly related to the down-regulation of VEGFA/PI3K/AKT pathway activity.更多还原