天然产物研究与开发 ›› 2021, Vol. 33 ›› Issue (9): 1558-1571.doi: 10.16333/j.1001-6880.2021.9.014

• 开发研究 • 上一篇    下一篇

吉祥草活性成分RCE-4抑制宫颈癌Ca Ski细胞增殖、侵袭和迁移的活性及机制研究

陈重旭1,游方芳1,金桂兰2,程凡1,邹坤1,邢翔飞2,奚炜2,陈剑锋1*   

  1. 1三峡大学生物与制药学院 三峡大学天然产物研究与利用湖北省重点实验室;2宜昌市第一人民医院,宜昌443002
  • 出版日期:2021-09-28 发布日期:2021-09-28
  • 基金资助:
    国家自然科学基金(81773952);湖北省卫生健康委员会中医药科研项目(ZY2019M028)

Study on the activity and mechanism of RCE-4,the active ingredient of Reineckia carnea,in inhibiting the proliferation,invasion and migration of cervical cancer Ca Ski cells

CHEN Chong-xu1,YOU Fang-fang1,JIN Gui-lan2, CHENG Fan1,ZOU Kun1,XING Xiang-fei2,XI Wei2,CHEN Jian-feng1*   

  1. 1Hubei Key Laboratory of Natural Products Research and Development,College of Biological and Pharmaceutical Sciences,China Three Gorges University;2The First People's Hospital of Yichang and The Peoples Hospital of China Three Gorges University,Yichang 443002,China

  • Online:2021-09-28 Published:2021-09-28

摘要:

本文探讨吉祥草活性单体RCE-4对宫颈癌Ca Ski细胞增殖、侵袭和迁移的影响及机制。采用MTT法和克隆形成实验检测RCE-4对Ca Ski细胞增殖的影响,划痕实验及Transwell小室实验研究RCE-4对Ca Ski细胞的迁移和侵袭的影响,间接免疫荧光实验和Western blot法检测RCE-4对β-catenin和YAP/TAZ蛋白入核转运的影响,采用Western blot法检测RCE-4对Wnt/β-catenin、Hippo/YAP信号路通及下游靶蛋白表达的影响。结果显示,RCE-4可以显著抑制Ca Ski细胞的增殖、迁移和侵袭;RCE-4处理后,β-catenin和YAP/TAZ蛋白入核减弱,GSK3β、p-β-catenin、p-YAP、p-LATS1的蛋白表达量增加,PAK4、p-GSK3ββ-catenin、MST1、LATS1、NUAK2、YAP/TAZ、YAP的蛋白表达量减少,下游靶蛋白MMPs、c-Myc、SOX2、CYR61、VCAM-1、ICAM-1的表达被抑制,提示RCE-4体外可能通过对Wnt/β-catenin和Hippo/YAP信号通路的调控发挥其抗Ca Ski细胞侵袭和迁移的作用。

关键词: RCE-4, 侵袭, 迁移, Wnt/β-catenin, Hippo/YAP

Abstract:

This article investigated the effect and mechanism of RCE-4 on proliferation,invasion and migration of cervical cancer Ca Ski cells.The effect of RCE-4 on the proliferation of Ca Ski cells was detected by MTT method and clone formation method.The scratch experiment and Transwell chamber experiment were performed to assess the effect of RCE-4 on the migration and invasion of Ca Ski cells.Indirect immunofluorescence experiment and Western blot method were used for detection.The effect of RCE-4 on the transport of β-catenin and YAP/TAZ proteins into the nucleus.Western blot was used to detect the effect of RCE-4 on the expression of Wnt/β-catenin,Hippo/YAP signaling pathway-related proteins and downstream target proteins.The results showed that RCE-4 can significantly inhibit the proliferation,migration and invasion of Ca Ski cells;after RCE-4 treatment,β-catenin and YAP/TAZ proteins enter the nucleus attenuated,and the protein expression of GSK3β,p-β-catenin,p-YAP,p-LATS1 increased,and the protein expression of PAK4, β-catenin,MST1,LATS1,NUAK2,YAP/TAZ,and YAP were decreased,and the expression of the downstream target proteins MMPs,c-Myc,SOX2,CYR61,VCAM-1,ICAM-1 were inhibited,suggesting that RCE-4 may exert its anti-Ca Ski cell invasion and migration effects through regulation of Wnt/β-catenin and Hippo/YAP signaling pathways in vitro.

Key words: RCE-4, invasion, metastasis, Wnt/β-catenin, Hippo/YAP 

中图分类号:  R966