Tripchlorolide activates p-ERK and induces autophagy in lung cancer A549 cells
10.3969/j.issn.1000-4718.2016.09.003
- VernacularTitle:雷公藤甲素通过 ERK 通路诱导肺癌 A549细胞自噬
- Author:
Wei WANG
;
Kun WANG
- Publication Type:Journal Article
- Keywords:
Lung cancer;
Tripchlorolide;
Cell proliferation;
Autophagy
- From:
Chinese Journal of Pathophysiology
2016;32(9):1551-1555
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the effects of tripchlorolide (TP) on proliferation and autophagy of human lung cancer A549 cells, and explore its mechanism.METHODS: MTT assay was performed to analyze the effect of TP on the viability of human lung cancer A549 cells.The A549 cells were treated with TP, and their autophagy was observed un-der the fluorescence microscope through acridine orange staining.Green fluorescence spots were observed by fluorescence microscopy through GFP-LC3 plasmid transfection experiment.The levels of LC3 and p-ERK in the A549 cells after TP treatment were determined by Western blot.RESULTS: The viability of human lung cancer A549 cells was significantly inhibited by TP in a dose-time dependent manner (P <0.05).The number of the intracellular acidic follicles dyed with bright red fluorescence was significantly increased after TP treatment in A549 cells.The number of green dot-like con-gregate autophagosomes in cell cytoplasm was significantly increased after TP treatment in the A549 cells transfected with GFP-LC3 plasmid, while the normal treatment only induced a few cells with autophagosome formation.At the same time, we did not observe the dot-like congregate autophagosomes after TP treatment in the A549 cells transfected with GFP-control plasmid.Compared with control group, the expression of LC3-II protein was up-regulated in A549 cells after TP treatment (P <0.01).Furthermore, treatment with TP in A549 cells for 48 h also led to a significant upregulation of phosphorylated form of ERK (P <0.01).In contrast, no significant change in the levels of total ERK protein was observed.Compared with 100 nmol/L TP group, TP +3-MA group down-regulated the protein levels of LC3-II (P <0.01) and p-ERK (P <0.01) in the A549 cells.CONCLUSION: TP significantly inhibits the growth of A549 lung cancer cells and induces the
autophagy, which may be correlated with upregulation of p-ERK protein.
