Research on the inhibitory effect of Tilianin on A549 cells and its mechanism
10.13699/j.cnki.1001-6821.2018.06.022
- VernacularTitle:田蓟苷对A549细胞的抑制作用及其机制研究
- Author:
Jian-Xia MENG
1
;
Ya-Li HUANG
;
Dong-Feng YIN
;
Xue-Bin WANG
;
Zhuo WANG
Author Information
1. 第二军医大学长海医院药学部
- Keywords:
Tilianin;
non-small cell lung cancer cell lines;
protein kinase B;
hypoxia inducible-factor-1α
- From:
The Chinese Journal of Clinical Pharmacology
2018;34(6):686-689
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the inhibitory effect of Tilianin on non -small cell lung cancer cell lines (A549) and the associated mechanisms.Methods The control group and 10,20,40,80 and 160 μmol· L-1 experimental group were treated with 0,10,20,40,80 and 160 μ mol· L-1Tilianin for 24 h,cell proliferation and toxicity test kit (CCK8) was used to observe the proliferation of A549 cells.The apoptosis of A549 cells in each group was detected by flow cytometry.In the hypoxia model group,A549 cells were cultured in hypoxic incubator for 4 h.In the hypoxia experimental group of 10,20 and 40 μmol · L-1Tilianin,A549 cells were pretreated with 10,20 and 40 μmol · L-1Tilianin for 4 h,and then cultured in hypoxic incubator for 4 h.Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the gene expressions of vascular endothelial growth factor(VEGF) and HIF-1α (HIF-1α).The protein expressions of VEGF-A,HIF-1α and p-Akt were detected by Western blot.Results Compared with control group,Tilianin significantly inhibited the proliferation of A549 in a dose -dependent manner.The inhibitory rates of proliferation of 10,20,40,80 and 160 μmol · L-1 experimental groups were (5.83 ±1.67)％,(6.77 ±0.87)％,(12.26 ±0.23)％,(22.97 ±0.50)％ and (46.24±1.44)％,the difference was statistically significant (P ＜0.05).The apoptosis rates of 10,20,40,80 and 160 μmol · L-1Tilianin were (6.80 ±0.62)％,(14.70 ±1.36)％,(24.76 ±4.37)％,(39.26±6.42)％ and (62.31 ±1.79)％,respectively,compared with the control group,the differences were statistically significant (P ＜ 0.05).In the hypoxia model group,the gene expression of HIF-1α was 4.30 ± 0.26,VEGF expression was 6.02 ± 0.53,compared with the control group,the differences was statistically significant (P ＜0.05).In 10,20,40 μmol · L-1Tilianin hypoxia experiment group,the gene expression of VEGF were 4.73 ±0.20,2.31 ±0.09 and 1.47 ±0.16,respectively.The expression of HIF-1α were 3.01 ± 0.11,1.81 ± 0.13 and 1.03 ± 0.16.Compared with hypoxia model group,the differences was statistically significant (P ＜ 0.05).The expression of p-AKT protein in hypoxic model group was (106.47 ± 2.08)％,the expression of HIF-1αwas (204.31 ± 8.35)％,the expression of VEGF-A was 212.30 ±4.80.The expression of p-AKT protein in 10,20,40 μmol · L-1 hypoxic experimental group were (87.51 ±2.72)％,(75.18 ± 1.67)％ and (32.40 ± 0.86)％,the protein expression levels of HIF-1α were (182.54 ± 6.42),(90.95 ± 2.76)％,(15.03 ± 4.21)％,the protein expression of VEGF-A were (156.38 ± 5.12) ％,(79.62 ± 2.84) ％ and (13.72 ± 4.64) ％,with significant difference (P ＜ 0.05).Conclusion Tilianin has the effect of inhibiting the proliferation and inducing apoptosis of A549 in vitro through AKT / HIF-1α signaling pathway.
