Dehydrocostus Lactone Inhibits Growth of Human Lung Cancer A549 Cells Through Activation of Apoptosis and Autophagy
10.13422/j.cnki.syfjx.20221626
- VernacularTitle:去氢木香内酯激活凋亡与自噬抑制人肺癌A549细胞生长
- Author:
Yingying TIAN
1
;
Yilin LI
1
;
Shiqiu TIAN
1
;
Hailuan PEI
1
;
Shangyue YU
1
;
Zijian WANG
2
;
Zeping ZUO
2
;
Zhibin WANG
1
Author Information
1. School of Chinese Materia Medica， Beijing University of Chinese Medicine， Beijing 100029， China
2. Research Institute of Beijing Tongrentang Co. Ltd.， Beijing 100079， China
- Publication Type:Journal Article
- Keywords:
dehydrocostus lactone;
lung cancer;
apoptosis;
autophagy;
protein kinase B （Akt）/mammalian target of rapamycin （mTOR）/signal transducer and activator of transcription 3 （STAT3） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2023;29(2):73-80
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo evaluate the effects of dehydrocostus lactone （DL） on the proliferation， apoptosis， and autophagy of human lung cancer cell A549 and to elucidate its related mechanism. MethodThe effect of DL with different concentrations （0， 5， 10， 15， 20， 25 μmol·L-1） on the proliferation of human lung cancer A549 cells was investigated by cell counting kit-8 （CCK-8）， and its impact on the clonogenic ability of A549 cells was studied by cell clonogenic assay. The concentrations 10， 20 μmol·L-1 were selected as DL low-dose group and high-dose group. Hoechst 33258 staining and western blot were used to observe the effect of DL on apoptosis of A549 cells. Autolysosomes were detected by acridine orange staining， and the expression level of microtubule-associated protein 1 light chain 3 （LC3） was determined by immunofluorescence and western blot. In addition， the effects of DL in combination with autophagy inhibitors bafilomycin A1 （BAF-A1） or 3-methyladenine （3-MA） on the autophagy of A549 cells was checked by CCK-8 assay. Finally， the role of DL in the regulation of A549 cell signaling pathway was explored by Western blot. ResultCompared with the conditions in the control group， the survival rate of A549 cells in the DL groups （10， 15， 20， 25 μmol·L-1） was decreased （P<0.01）， and 5 μmol·L-1 DL could inhibited the formation of A549 clone cells （P<0.01）， indicating that DL could inhibit the proliferation of human lung cancer A549 cells. The number of apoptotic cells was higher in both DL low-dose and high-dose groups than that in the control group， and the expression of apoptosis-related proteins poly （ADP ribose） polymerase （PARP） and B lymphocytoma-2 （Bcl-2）-associated X protein （Bax） were up-regulated （P<0.05， P<0.01）， while the expression of Bcl-2 was down-regulated （P<0.01） in DL high-dose group. The acridine orange staining showed that the orange fluorescence in the DL high-dose group was enhanced compared with that in the control group， indicating that DL could dramatically promote the formation of autolysosomes. Moreover， 20 μmol·L-1 DL could increase the orange fluorescent particles of LC3 and up-regulated the expression level of LC3 Ⅱ （P<0.01）. After addition of autophagy inhibitors， the sensitivity of A549 cells to the effects of DL was attenuated （P<0.01）， which suggested that autophagy was involved in DL-induced A549 cell death. Compared with the control group， DL high-dose group had increased expression of autophagy-related protein 3 （Atg3） and autophagy-related protein 5 （Atg5） while reduced phosphorylation levels of protein kinase B （Akt）， mammalian target of rapamycin （mTOR） and signal transducer and activator of transcription 3 （STAT3） （P<0.05， P<0.01）. ConclusionDL could activate apoptosis and autophagy to inhibit the proliferation and clonogenic ability of A549 cells via suppressing Akt/mTOR/STAT3 signaling pathway.
