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山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (1): 10-16.doi: 10.6040/j.issn.1671-7554.0.2022.0885

• 基础医学 • 上一篇    

黄芪皂苷Ⅱ对肾透明细胞癌细胞生长抑制作用及机制

赵凯1,尹心宝2,张宗亮2,王振林2,朱冠群2,王科2   

  1. 1.辽宁中医药大学, 辽宁 沈阳 116600;2.青岛大学附属医院泌尿外科, 山东 青岛 266003
  • 发布日期:2023-01-10
  • 通讯作者: 王科. E-mail:wangke6910@sina.com
  • 基金资助:
    国家自然科学基金(31971191)

Inhibitory effect and mechanism of astragaloside Ⅱ on renal clear cell carcinoma cells

ZHAO Kai1, YIN Xinbao2, ZHANG Zongliang2, WANG Zhenlin2, ZHU Guanqun2, WANG Ke2   

  1. 1. Liaoning University of Traditional Chinese Medicine, Shenyang 116600, Liaoning, China;
    2. Department of Urology, Affiliated Hospital of Qingdao University, Qingdao 266003, Shandong, China
  • Published:2023-01-10

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摘要： 目的 探讨黄芪皂苷Ⅱ(As Ⅱ)对肾透明细胞癌786-O细胞增殖的影响及潜在机制。 方法 噻唑兰(MTT)法和克隆实验检测As Ⅱ对786-O细胞的生长和集落形成的抑制作用。细胞流式术检测AS Ⅱ的凋亡诱导作用。随后,用免疫印迹法检测不同处理组细胞PI3K-AKT-mTOR通路蛋白及凋亡执行蛋白的变化,最后利用PI3K-AKT-mTOR通路激活剂IGF-1与As Ⅱ对细胞共处理后,流式细胞术检测细胞凋亡水平的变化,进一步验证PI3K-AKT-mTOR通路在凋亡诱导中的作用。 结果 As Ⅱ对786-O细胞的增殖抑制效应具有浓度和时间依赖性,时间与浓度之间有交互作用(F时间=513.00, P<0.001;F浓度=1 678.00, P<0.001;F时间×浓度=18.23, P<0.001),并可抑制细胞集落形成。As Ⅱ可诱导细胞凋亡并诱导凋亡执行蛋白cleved caspase-3的表达(P0 μmol/L As Ⅱ组 vs 10 μmol/L As Ⅱ组= 0.013 9,P0 μmol/L As Ⅱ组 vs 20 μmol/L As Ⅱ组<0.001)。As Ⅱ可抑制PI3K、AKT、mTOR蛋白的磷酸化(p-PI3K: P0 μmol/L As Ⅱ组 vs 20 μmol/L As Ⅱ组=0.032 4;p-mTOR: P0 μmol/L As Ⅱ组 vs 10 μmol/L As Ⅱ组=0.004 1,P0 μmol/L As Ⅱ组 vs 20 μmol/L As Ⅱ组<0.001;p-AKT:P0 μmol/L As Ⅱ组 vs 10 μmol/L As Ⅱ组=0.003 2,P0 μmol/L As Ⅱ组 vs 20 μmol/L As Ⅱ组=0.001 2)。IGF-1可以逆转As Ⅱ对AKT的磷酸化(P=0.006 8),并减弱AsⅡ对786-O细胞的凋亡诱导效应。 结论 As Ⅱ可抑制肾癌780-O细胞的增殖和集落形成,并通过抑制PI3K-AKT-mTOR信号通路的磷酸化,上调cleved caspase-3的表达,进而诱导细胞凋亡。

关键词: 肾透明细胞癌, 黄芪皂苷Ⅱ, 肿瘤治疗, 凋亡

Abstract: Objective To investigate the effect of astragaloside Ⅱ(As Ⅱ)on the proliferation of renal clear cell carcinoma 786-O cells and its potential mechanism. Methods The inhibitory effect of As Ⅱ on the growth and colony formation of 786-O cells was detected by MTT assay and cloning assay. Cell flow cytometry was used to detect the apoptosis-inducing effect of AS Ⅱ. Subsequently, Western blotting was used to detect the changes of PI3K-AKT-mTOR pathway protein and apoptosis executioner protein in cells treated with different concentrations of As Ⅱ. After co-treatment of cells with PI3K-AKT-mTOR pathway activator IGF-1 and As Ⅱ, flow cytometry was used to detect the changes of apoptosis and further verify the role of PI3K-AKT-mTOR pathway in apoptosis induction. Results As Ⅱ inhibited the proliferation of 786-O cells in a concentration- and time-dependent manner, with an interaction between time and concentration(Ftime=513.00, P<0.001; Fconcentration=1 678.00, P<0.001; Ftime×concentration=18.23, P<0.001), and inhibited cell colony formation. As Ⅱ induced apoptosis and increased the expression of cleved caspase-3, an apoptotic executioner protein(P0 μmol/L As Ⅱ group vs 10 μmol/L As Ⅱ group=0.013 9,P0 μmol/L As Ⅱ group vs 20 μmol/L As Ⅱ group<0.001). As Ⅱ inhibited the phosphorylation of PI3K, AKT and mTOR proteins(p-PI3K: P0 μmol/L As Ⅱ group vs 20 μmol/L As Ⅱ group=0.032 4;p-mTOR: P0 μmol/L As Ⅱ group vs 10 μmol/L As Ⅱ group=0.004 1,P0 μmol/L As Ⅱ group vs 20 μmol/L As Ⅱ group<0.001;p-AKT:P0 μmol/L As Ⅱ group vs 10 μmol/L As Ⅱ group=0.003 2,P0 μmol/L As Ⅱ group vs 20 μmol/L As Ⅱ group=0.001 2). IGF-1 reversed the phosphorylation of AKT by As Ⅱ(P=0.006 8), and attenuated the apoptosis-inducing effect of As Ⅱ on 786-O cells. Conclusion As Ⅱ can inhibit the proliferation and colony formation of renal carcinoma 780-O cells, and up-regulate the expression of cleved caspase-3 by inhibiting the phosphorylation of PI3K-AKT-mTOR signaling pathway, which in turn induces apoptosis.

Key words: Clear cell renal carcinoma, Astragaloside Ⅱ, Cancer therapy, Apoptosis

中图分类号: 

  • R737.11
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