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  

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

作　　者：刘倩 胡春萍 曹鹏[1,2] 方志军 孙晓艳[1,2] 

机构地区：[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　　处：《天然产物研究与开发》2017年第12期2030-2035,共6页Natural Product Research and Development

基　　金：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)

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。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, MTr 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-apop- totic protein Bcl-2,and up-regulated expression of apoptotic protein Bax in ArBu-treated cells. These results demonstra- ted 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.

关 键 词：沙蟾毒精 表皮生长因子受体 肺癌 细胞凋亡 

分 类 号：Q939.9[生物学—微生物学]