实用医学杂志 ›› 2020, Vol. 36 ›› Issue (23): 3199-3205.doi: 10.3969/j.issn.1006⁃5725.2020.23.007

• 基础研究 • 上一篇    下一篇

芍药苷抑制Src/STAT3信号通路协同替莫唑胺抑制胶质瘤细胞增殖和迁移

高卓维, 陈广鸿, 符路娣   

  1. 1广州中医药大学顺德医院(广东佛山528300);2 南方医科大学中医药学院分子生物学实验室国家中医药管理局三级科研实验室(广州510515);3 广州中医药大学实验动物中心(广州510405)

  • 出版日期:2020-12-10 发布日期:2020-12-23
  • 通讯作者: 符路娣E⁃mail:fuludi123@163.com
  • 基金资助:
    国家自然科学基金(编号:81703914)

Paeoniflorin inhibits Src/STAT3 signaling pathway and temozolomide inhibits proliferation and migration of primary human glioma cells

GAO Zhuowei,CHEN Guanghong,FU Ludi   

  1. Shunde Hospital,Guangzhou University of Traditional Chinese Medicine,Foshan 528300,China
  • Online:2020-12-10 Published:2020-12-23
  • Contact: FU Ludi E⁃mail:fuludi123@163.com

摘要:

目的 观察芍药苷协同替莫唑胺对原代胶质瘤细胞体外增殖和迁移的影响,初步探讨其作用机制。方法 MTS 法检测芍药苷、替莫唑胺以及两药联合对原代胶质瘤胶质瘤细胞活力的影响,应用等效线图分析协同效应。采用EdU 染色检测细胞增殖,Transwell 细胞迁移实验检测细胞迁移,通过反向药效团对接技术探索芍药苷抑制原代胶质瘤细胞的潜在作用靶点并采用Western blot 进行验证。结果 芍药苷及替莫唑胺呈浓度和时间依赖性抑制原代胶质瘤细胞活力,两药联合具有协同抑制作用;两药联合对胶质瘤细胞增殖和迁移的抑制作用明显高于芍药苷及替莫唑胺单药组;反向药效团对接技术提示Src 可能是芍药苷的作用靶点,Western blot 检测表明芍药苷能够显著抑制p⁃Src 和p⁃STAT3 的表达,芍药苷通过抑制Src/STAT3 信号通路协同替莫唑胺增强了对增殖和迁移相关蛋白的抑制作用。结论 芍药苷能显著提高替莫唑胺的抗胶质瘤作用,此协同作用主要通过抑制Src/STAT3 信号通路的激活,从而进一步增强对增殖和迁移相关蛋白的抑制作用,抑制胶质瘤细胞增殖和迁移。

关键词: 胶质瘤, 替莫唑胺, 芍药苷, Src/STAT3信号通路, 增殖, 迁移

Abstract:

Objective To observe the combinative effects of paeoniflorin and temozolomide on the prolifera⁃tion and migration of primary human glioma cells in vitro,and preliminary explore its mechanism of action. Methods The MTS method was used to detect the effects of paeoniflorin,temozolomide and the combination of the two drugson the viability of primary human glioma cells. The equivalent line diagram was used to analyze the synergisticeffect. EdU staining was used to detect cell proliferation. Transwell cell migration experiment was used to detect cellmigration. The reverse pharmacophore docking technique was used to explore the potential target of paeoniflorin toinhibit glioma cells and Western blot to verify. Results Paeoniflorin and temozolomide inhibited primary humanglioma cells viability in a concentration⁃ and time⁃dependent manner. The combination of the two drugs had a syner⁃gistic inhibitory effect;the inhibitory effect of the combination of the two drugs on cell proliferation and migration wassignificantly higher than that of the paeoniflorin and temozolomide monotherapy group. The effect group dockingtechnique suggests that Src may be the target of paeoniflorin. Western blot analysis showed that paeoniflorin cansignificantly inhibit the expression of p⁃Src and p⁃STAT3. Inhibition of migration⁃related proteins. Conclusion Paeoniflorin can significantly improve the anti ⁃ glioma effect of temozolomide. This synergistic effect is mainlythrough inhibiting the activation of Src/STAT3 signaling pathway,thereby further enhancing the inhibitory effect onproliferation and migration⁃related proteins and inhibiting the proliferation and migration of glioma cells.

Key words: glioma, temozolomide, paeoniflorin, Src/STAT3 signaling pathway, proliferation, migration