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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
[1] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020[J]. CA Cancer J Clin, 2020, 70(1): 7-30.
[2] Rysz J, Franczyk B, Ławiński J, et al. Characteristics of clear cell papillary renal cell carcinoma(ccpRCC)[J]. Int J Mol Sci, 2021, 23(1): 151.
[3] Canil C, Hotte S, Mayhew LA, et al. Interferon-alfa in the treatment of patients with inoperable locally advanced or metastatic renal cell carcinoma: a systematic review [J]. Can Urol Assoc J, 2010, 4(3): 201-208.
[4] Nielsen OH, Grimm D, Wehland M, et al. Anti-angiogenicdrugs in the treatment of metastatic renal cell carcinoma: advances in clinical application[J]. Curr Vasc Pharmacol, 2015, 13(3): 381-391.
[5] Stone L. Kidney cancer: no advantage of adjuvant sunitinib or sorafenib [J]. Nat Rev Urol, 2016, 13(5): 240-241.
[6] 周岱翰. 中医肿瘤学发展历程的反思与展望[J]. 中医肿瘤学杂志, 2019, 1(1): 1-5, 9. ZHOU Daihan. Reflection and prospect of traditional Chinese medicine oncology evelopment [J]. Journal of Oncology in Chinese Medicine, 2019, 1(1): 1-5, 9.
[7] 张隽开,王忠裕,丁大朋,等. 黄芪对肝癌耐药细胞株Bel/Fu化疗敏感性的影响[J]. 中国中西医结合外科杂志, 2008, 14(4): 398-402. ZHANG Junkai, WANG Zhongyu, DING Dapeng, et al. Effect of astragulus injection on chemosensitivity of drug-resistant hepatic carcinoma Bel/Fu cell line [J]. Chinese Journal of Surgery of Integrated Traditional and Western Medicine, 2008, 14(4): 398-402.
[8] 王培培. 黄芪皂苷Ⅱ和Ⅳ对人肝癌细胞BEL-7402/5-FU的耐药逆转作用及其机制研究[D]. 合肥:安徽医科大学, 2010.
[9] 谢荣丹,龙凤. 黄芪及黄芪有效成分抗乳腺癌作用的研究[J]. 世界最新医学信息文摘, 2018, 18(92): 67-68. XIE Rongdan, LONG Feng. The study on the effect of astragalus and its active components against breast cancer [J]. World Latest Medicine Information(Electronic Version), 2018, 18(92): 67-68.
[10] 熊斌, 鲁伟. 转移性肾癌治疗的现状和进展[J]. 中华泌尿外科杂志, 2021, 42(4): 308-311.
[11] 徐可, 周启东. 晚期肾癌治疗与基础研究进展[J]. 上海医学, 2019, 42(3): 143-146.
[12] 王媛, 吴英, 董丽,等. 转移性肾癌的治疗新进展[J]. 临床误诊误治, 2018, 31(2): 114-116.
[13] Wang YP, Li XY, Song CQ, et al. Effect of astragaloside IV on T, B lymphpcyte proliferation and peritoneal macrophage function in mice[J]. Acta Pharmacol Sin, 2002, 23(3): 263-266.
[14] Qi Y, Gao F, Hou L, et al. Anti-inflammatory and immunostimulatory activities of astragalosides[J]. Am J Chin Med, 2017, 45(6): 1157-1167.
[15] 高崇婷, 桂定坤, 汪年松,等. 黄芪皂苷Ⅱ对STZ诱导的糖尿病大鼠的肾脏保护作用研究[J]. 现代生物医学进展, 2020, 20(23): 4401-4406, 4417. GAO Chongting, GUI Dingkun, WANG Niansong, et al. Renal protective effect of astragaloside-Ⅱ on STZ-induced diabetic rats [J]. Progress in Modern Biomedicine, 2020, 20(23): 4401-4406, 4417.
[16] 武超,许杜娟,杨翠,等. 黄芪皂苷Ⅱ增加5-氟尿嘧啶对人肝癌细胞株HepG2增殖抑制作用[J]. 安徽医科大学学报, 2016, 51(1): 78-82. WU Chao, XU Dujuan, YANG Cui, et al. Increasing inhibition of human hepatic cancer HepG2 cells to 5-fluorouracil by Astragaloside II [J]. Acta Universitatis Medicinalis Anhui, 2016, 51(1): 78-82.
[17] 王敏, 郑喜, 祁燕,等. 黄芪皂苷II抗肝癌肺转移效应及作用机理的研究[J]. 中药药理与临床, 2019, 35(6): 41-45. WANG Min, ZHENG Xi, QI Yan, et al. Effect and mechanism of astragaloside II on pulmonary metastatic of hepatoma in mice [J]. Pharmacology and Clinics of Chinese Materia Medica, 2019, 35(6): 41-45.
[18] Guo H, German P, Bai SS, et al. The PI3K/AKT pathway and renal cell carcinoma [J]. J Genet Genomics, 2015, 42(7): 343-353.
[19] 马建辉. 肾癌细胞mTOR信号通路异常与依维莫司作用机制[J]. 中华泌尿外科杂志, 2013, 34(3): 229-231.
[20] 宿恒川, 孙福康. PI3K/Akt/mTOR信号通路及mTOR抑制剂在泌尿系统肿瘤中的研究进展[J]. 上海交通大学学报(医学版), 2012, 32(5): 674-678. SU Hengchuan, SUN Fukang. Research progress of PI3K/Akt/mTOR signaling pathway and m TOR inhibitors in urologic malignancies [J]. Journal of Shanghai Jiaotong University(Medical Science), 2012, 32(5): 674-678.
[21] 李文超,陈军宝,卢宏柱. PI3K/Akt/mTOR信号通路在肾细胞癌中的研究进展[J]. 医学综述, 2014, 20(16): 2930-2932. LI Wenchao, CHEN Junbao, LU Hongzhu. The research progress of PI3K/Akt/mTOR signaling pathway in renal cell carcinoma [J]. Medical Recapotulate, 2014, 20(16): 2930-2932.
[22] 王娟. 冬青素A经PI3K/AKT/mTOR通路抑制血管新生抗肝癌增殖[D]. 扬州:扬州大学, 2016.
[23] 史颖弘,樊嘉,周俭,等. PI3K/AKT/mTOR信号通路在肝癌细胞自体吞噬中的作用研究[J]. 中华消化外科杂志, 2007, 6(1): 39-43.
[24] Guo C, Gasparian AV, Zhuang Z, et al. 9-Aminoacridine-based anticancer drugs target the PI3K/AKT/mTOR, NF-kappa B and p53 pathways[J]. Oncogene, 2009, 28(8): 1151-1161.
[25] Fuereder T, Jaeger-Lansky A, Hoeflmayer D, et al. mTOR inhibition by everolimus counteracts VEGF induction by sunitinib and improves anti-tumor activity against gastric cancer in vivo[J]. Cancer Lett, 2010, 296(2): 249-256.
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