天然产物研究与开发 ›› 2017, Vol. 29 ›› Issue (1): 135-140.doi: 10.16333/j.1001-6880.2017.1.026

• 开发研究 • 上一篇    下一篇

柠檬酚对人肝癌HepG2细胞增殖的影响

刘瑛,陆国寿*,卢文杰,王丽,黄周锋   

  1. 广西中药质量标准研究重点实验室广西壮族自治区中医药研究院,南宁 530022
  • 出版日期:2017-01-31 发布日期:2017-02-17

Effect of Citrusinol on Proliferation of Human Hepatocellular Carcinoma Cells HepG2

LIU Ying,LU Guo-shou*,LU Wen-jie,WANG Li,HUANG Zhou-feng   

  1. Guangxi Key Laboratory of Traditional Chinese Medicine Quality StandardsGuangxi Institute of Traditional Medical and Pharmaceutical Sciences,Nanning 530022,China
  • Online:2017-01-31 Published:2017-02-17

摘要: 为了探讨柠檬酚对人肝癌HepG2细胞增殖的影响,本文利用四甲基偶氮唑蓝(MTT)法、细胞生长曲线以及细胞克隆形成实验检测了柠檬酚对细胞生长和克隆形成的影响,并应用HE、吖啶橙染色、流式细胞术以及激光扫描共聚焦显微镜分别观察和检测了不同浓度柠檬酚作用后细胞形态、细胞周期和细胞骨架蛋白F-actin的变化。结果显示,柠檬酚能明显抑制人HepG2细胞生长,且呈浓度依赖性,其IC50值为79.14 μmol/L,柠檬酚给药组细胞克隆率明显明显低于空白对照组;不同浓度的柠檬酚在作用HepG2细胞24h后,细胞均出现了明显的皱缩、胞浆空泡化和细胞核固缩的形态学改变;同时,70 μmol/L和80 μmol/L的柠檬酚处理组G2/M期细胞比例显著高于对照组(22.72±1.07,28.68±1.36 vs 16.34±0.66,P<0.01);而共聚焦显微镜观察发现,柠檬酚能引起HepG2细胞骨架蛋白F-actin的解聚和断裂。综上,推测柠檬酚可能是通过破坏细胞的Actin骨架,阻止细胞有丝分裂,细胞周期阻滞在G2/M期,从而抑制人肝癌HepG2细胞的增殖。

关键词: 柠檬酚, HepG2细胞, 增殖抑制, 细胞周期, 细胞骨架蛋白

Abstract: In order to study the effect of citrusinol on human hepatocellular carcinoma cells HepG2,terazolium salt colorimetry assay(MTT ),cell growth curve and colony formation assay was used to detected the proliferation and colony formation.The morphology of cells was observed by HE staining and Acridine orange staining.Cell cycle changes were detected by Flow Cytometry(FCM).The morphological changes of cytoskeletal protein F-actin were observed by laser confocal microscopy.The results showed thatcitrusinol inhibited HepG2cells proliferation and colony formation in a dose-dependent manner.IC50 of citrusinol was 79.14 μmol/L after 24 hours.The morphology of HepG2 cells was changed,showing cell shrinkage,cytoplasm vacuoles and nuclear chromatin pyknosis.The percentage of G2/M phase of HepG2 cells treated with 70,80 μmol/L citrusinol was significantly higher than that of the control group (22.72±1.07,28.68±1.36 vs 16.34±0.66,P<0.01).F-actin cytoskeletal protein was showed from polymerization to depolymerization after citrusinol treatment.Based on the experimental results,it was concluded that citrusinol can prevent cell mitosis to arrest the cell cycle in G2/M phase,by disrupting Actin cytoskeleton,which may be one of the mechanisms of citrusinol's anti-tumor effect.

Key words: citrusinol, HepG2 cell, antiproliferation, cell cycle, cytoskeletal protein

中图分类号: 

R963