黄芪甲苷对舌鳞癌细胞增殖、迁移、侵袭、凋亡的作用及机制

何泽禹; 朱永娜; 姜丽娜; 张晓筱; 刘茜

doi:10.13898/j.cnki.issn.2097-5252.2025.04.004

黄芪甲苷对舌鳞癌细胞增殖、迁移、侵袭、凋亡的作用及机制

Role of astragaloside Ⅳ in proliferation,migration,invasion, and apoptosis of tongue squamous cell carcinoma cells and its mechanism

摘要

摘要: 目的：探讨黄芪甲苷(AS-Ⅳ)对人舌鳞状细胞癌CAL-27细胞增殖、迁移、侵袭、凋亡的影响及其机制。方法：体外培养CAL-27细胞。不同浓度AS-Ⅳ(0、5、10、20、40、80、160 μmol/L)处理CAL-27细胞24、48、72 h，采用CCK-8法检测细胞增殖情况。0、20、40、80 μmol/L AS-Ⅳ处理CAL-27细胞24 h，采用Transwell实验检测细胞迁移和侵袭能力，流式细胞术检测细胞凋亡情况。将CAL-27细胞分为对照组、转化生长因子-β1(TGF-β1)组(10 ng/mL TGF-β1)、联合用药组(40 μmol/L AS-Ⅳ+10 ng/mL TGF-β1)，Transwell实验检测各组细胞迁移和侵袭能力，Western blotting分析转化生长因子-βⅡ受体(TGF-β RⅡ)、Smad4蛋白表达水平。结果：CCK-8结果显示，与0 μmol/L AS-Ⅳ组比较，20、40、80 μmol/L AS-Ⅳ作用24、48、72 h吸光度值均降低(P<0.05~P<0.01)。流式细胞术结果显示，与0 μmol/L AS-Ⅳ组比较，20、40、80 μmol/L AS-Ⅳ作用24 h均能明显诱导CAL-27细胞凋亡(P<0.01)；与20、40 μmol/L AS-Ⅳ组比较，80 μmol/L AS-Ⅳ组细胞凋亡率明显增加(P<0.01)。Transwell实验结果显示，与0 μmol/L AS-Ⅳ组相比，20、40、80 μmol/L AS-Ⅳ均能明显抑制CAL-27细胞迁移和侵袭能力(P<0.01)，且随着AS-Ⅳ浓度的增加，迁移和侵袭细胞数逐渐减少(P<0.01)；转化生长因子-β1(TGF-β1)组细胞迁移和侵袭数明显高于对照组(P<0.01)，联合组细胞迁移和侵袭数高于对照组(P<0.05和P<0.01)，但低于TGF-β1组(P<0.05)。Western blotting结果显示，TGF-β1组与联合组的TGF-β RⅡ和Smad4蛋白水平均明显高于对照组(P<0.01)，联合组的TGF-β RⅡ和Smad4蛋白水平均明显低于TGF-β1 组(P<0.01)。结论：AS-Ⅳ可以抑制舌鳞状细胞癌CAL-27细胞增殖、迁移、侵袭，诱导细胞凋亡。AS-Ⅳ可能通过阻断TGF-β1介导的TGF-β/Smad信号通路调节CAL-27细胞的迁移和侵袭能力。

Abstract: Objective: To investigate the effects of astragaloside Ⅳ (AS-Ⅳ) on the proliferation,migration,invasion,and apoptosis of human tongue squamous cell carcinoma CAL-27 cells and its mechanism. Methods: The CAL-27 cells were cultured in vitro.The cells were treated with different concentrations of AS-Ⅳ (0, 5, 10, 20, 40, 80, 160 μmol/L) for 24,48,72 hours,and cell proliferation was detected by CCK-8 method.After treatment with 0,20,40,and 80 μmol/L AS-Ⅳ for 24 hours,cell migration and invasion abilities of CAL-27 cells were evaluated by Transwell assay,and apoptosis was analyzed by flow cytometry.The CAL-27 cells were divided into the control group,transforming growth factor-β1 (TGF-β1) group (10 ng/mL TGF-β1), and combination group (40 μmol/L AS-Ⅳ+10 ng/mL TGF-β1),and the cell migration and invasion abilities of each group were detected by Transwell assay,the protein expression levels of transforming growth factor-β receptor Ⅱ (TGF-β RⅡ) and Smad4 were analyzed by Western blotting. Results: The results of CCK-8 method showed that compared with the 0 μmol/L AS-Ⅳ group,the optical density values of 20,40,and 80 μmol/L AS-Ⅳ treatment for 24,48,and 72 hours were reduced (P<0.05 to P<0.01).The results of flow cytometry indicated that compared with the 0 μmol/L AS-Ⅳ group,20,40,and 80 μmol/L AS-Ⅳ treatment for 24 hours could significantly induce apoptosis in CAL-27 cells (P<0.01);compared with the 20 μmol/L AS-Ⅳ group and 40 μmol/L AS-Ⅳ group,the apoptosis rate in the 80 μmol/L AS-Ⅳ group was significantly increased (P<0.01).The results of Transwell assay found that compared with the 0 μmol/L AS-Ⅳ group,20,40,and 80 μmol/L AS-Ⅳ could significantly inhibit the migration and invasion abilities of CAL-27 cells (P<0.01),and the number of migrating and invading cells gradually decreased with the increase of AS-Ⅳ concentration (P<0.01);the number of migrating and invading cells in the transforming growth factor-β1 (TGF-β1) group was significantly higher than that in the control group (P<0.01),the number of migrating and invading cells in the combination group was higher than that in the control group (P<0.05 and P<0.01),but lower than that in the TGF-β1 group (P<0.05).The results of Western blotting implied that the levels of TGF-β RⅡ and Smad4 proteins in the TGF-β1 group and combination group were significantly higher than those in the control group (P<0.01),while the levels of TGF-β RⅡ and Smad4 proteins in the combination group were significantly lower than those in the TGF-β1 group (P<0.01). Conclusions: AS-Ⅳ can induce apoptosis and inhibit proliferation,migration,and invasion of tongue squamous cell carcinoma CAL-27 cells.AS-Ⅳ may regulate the migration and invasion abilities of CAL-27 cells by blocking the TGF-β/Smad signaling pathway mediated by TGF-β1.

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