天然产物研究与开发 ›› 2017, Vol. 29 ›› Issue (12): 2030-2035.doi: 10.16333/j.1001-6880.2017.12.006

• 研究论文 • 上一篇    下一篇

沙蟾毒精诱导肺癌PC-9细胞的凋亡效应及其作用机制初探

刘倩1,2,胡春萍1,2,曹鹏1,2,方志军1,2*,孙晓艳1,2*   

  1. 1南京中医药大学 附属中西医结合医院;2江苏省中医药研究院 细胞与分子生物学实验室,南京 210028
  • 出版日期:2018-01-04 发布日期:2018-01-09
  • 基金资助:

    公益性行业科研专项经费项目(201507002);江苏省省属公益院所科研条件与能力建设项目(BM2015026)

The Effects and Mechanisms of Apoptosis Induced by Arenobufagin in Lung Cancer PC-9 Cells

LIU Qian1,2,HU Chun-ping1,2,CAO Peng1,2,FANG Zhi-jun1,2*,SUN Xiao-yan1,2*   

  1. 1Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine, 2Laboratory of Cellular and Molecular Biology,Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing 210028,Jiangsu,China
  • Online:2018-01-04 Published:2018-01-09

摘要: 本文主要研究沙蟾毒精(Arenobufagin,ArBu)对非小细胞肺癌PC-9细胞的凋亡作用及其作用机制。以非小细胞肺癌PC-9细胞为研究对象,采用MTT法检测ArBu对肺癌PC-9细胞的生长抑制作用,结果发现ArBu各浓度组在不同时间点均对PC-9细胞具有显著的抑制作用。荧光显微镜观察经Hoechst 33258染色的细胞形态,结果显示ArBu给药组细胞减少且出现了染色质固缩、核碎裂、凋亡小体形成,说明ArBu诱导了细胞凋亡。进一步,流式细胞检测仪结果显示细胞凋亡呈明显的量效关系。此外,Western blot结果显示ArBu抑制EGFR/RAF/MEK/ERK信号通路蛋白的磷酸化,下调抗凋亡蛋白Bcl-2的表达,上调促凋亡蛋白Bax的表达。上述实验结果表明,ArBu对非小细胞肺癌细胞系PC-9具有明显的生长抑制和促凋亡作用,可作为抗非小细胞肺癌的先导化合物进一步研究。

关键词: 沙蟾毒精, 表皮生长因子受体, 肺癌, 细胞凋亡

Abstract: This work mainly investigated the effects of Arenobufagin (ArBu) on the viability and apoptosis of non-small cell lung cancer (NSCLC) cells and explored the mechanisms of ArBu-induced apoptosis.Taken NSCLC cells PC-9 as a study model,MTT assay was performed to detect the inhibitory effects of ArBu on PC-9.The results showed that ArBu significantly suppressed the growth of PC-9 cells at various concentrations and treating times.Hoechst 33258 staining demonstrated that ArBu -treatment reduced the cell number,caused chromatin condensation,nucleic fragmentation and apoptotic body formation,indicating the induction of apoptosis.Furthermore,the results of flow cytometry showed that a significant quantitative relationship between apoptosis and the concentrations of ArBu.In addition,western blot detected decreased phosphorylation of EGFR/RAF/MEK/ERK signal pathway proteins,down-regulated expression of anti-apoptotic protein Bcl-2,and up-regulated expression of apoptotic protein Bax in ArBu-treated cells.These results demonstrated that ArBu can significantly decrease the viability and induce apoptosis of PC-9 cells,suggesting that ArBu has the potential to be further studied as a leader compound for the treatment of lung cancer.

Key words: Arenobufagin, EGFR, lung cancer, apoptosis

中图分类号: 

Q939.9