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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