Effect of Yiqi Fuzheng Jiedu Decoction on Autophagy and Growth of A549 Cells： An Exploration Based on p53/AMPK Signaling Pathway
10.13422/j.cnki.syfjx.20212121
- VernacularTitle:基于p53/AMPK信号通路观察益气扶正解毒汤对A549细胞自噬水平和生长的影响
- Author:
Qiao-lan WU
1
;
Ting SONG
2
;
Ze-tao CHEN
2
;
Xiu-bao CHEN
2
;
Yi-chen ZHANG
1
;
Wei-da CHEN
2
Author Information
1. First Clinical Medical College， Shandong University of Traditional Chinese Medicine（TCM）， Jinan 250014， China
2. Affiliated Hospital of Shandong University of TCM， Jinan 250014， China
- Publication Type:Research Article
- Keywords:
Yiqi Fuzheng Jiedu decoction （YQFZJDD）;
A549 cells;
autophagy;
p53/adenosine monophosphate-activated protein kinase （AMPK） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(22):65-75
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of Yiqi Fuzheng Jiedu decoction （YQFZJDD） on autophagy and growth of A549 cells. Method:A549 cells were intervened with YQFZJDD-containing serum prepared in advance. The levels of microtubule-associated protein 1 light chain 3 （LC3）， homologue of yeast autophagy-related gene 6 （Beclin1）， p62， p53， adenosine 5'-monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mammalian target of rapamycin （mTOR）， and phosphorylated mTOR （p-mTOR） were detected by Western blot assay， and microtubule-associated protein 1A/1B light chain 3B （MAP1LC3B） by immunofluorescence （IF） assay. The proliferation， invasion， and senescence of A549 cells were separately measured by 5-ethynyl-2'-deoxyuridine （EDU） staining， Transwell assay， and β-galactosidase staining. In the presence of autophagy inhibitor 3-methyladenine （3-MA， 5 mmol·L-1）， the cells were divided into the 10% fetal bovine serum （blank） group， 10% control serum （control） group， low-， medium-， and high-dose 10% YQFZJDD-containing serum groups， and high-dose 10% YQFZJDD-containing serum + 3-MA group， followed by the measurement of A549 cell proliferation， invasion， and senescence. In the adoption of p53 inhibitor Pifithrin-α （PFT-α， 10 μmol·L-1）， the cells were divided into the control group， PFT-α group， high-dose YQFZJDD-containing serum group， and high-dose YQFZJDD-containing serum + PFT-α group. Then the LC3-Ⅱ and Beclin1 expression， MAP1LC3B fluorescence intensity， as well as A549 cell proliferation， invasion and senescence were determined. Result:Compared with the blank group and control group， YQFZJDD-containing serum at the medium and high doses up-regulated the protein expression levels of LC3-Ⅱ and Beclin1 in A549 cells after 48-h intervention in a dose-dependent manner （P<0.01）. Besides， YQFZJDD-containing serum at the low-， medium-， and high-doses down-regulated p62 and p-mTOR/mTOR expression （P<0.05， P<0.01）， elevated p53 and p-AMPK/AMPK （P<0.01）， decreased the number of proliferative and invasive cells， increased the number of senescent cells （P<0.01）， and enhanced the IF intensity of MAP1LC3B， with the optimal effect observed in the high-dose YQFZJDD-containing serum group （P<0.01）. Compared with the high-dose YQFZJDD-containing serum group， the high-dose YQFZJDD-containing serum + 3-MA group and high-dose YQFZJDD-containing serum + PFT-α group exhibited significantly increased proliferative and invasive cells but decreased senescent cells （P<0.05， P<0.01）. Meanwhile， the IF intensity of MAP1LC3B in the high-dose YQFZJDD-containing serum + PFT-α group was weakened and the LC3-Ⅱ and Beclin1 protein expression levels declined （P<0.05， P<0.01）. Conclusion:YQFZJDD promotes the autophagy of A549 cells through the p53/AMPK signaling pathway to inhibit their proliferation， invasion and migration but accelerate senescence， thus playing a crucial role in inhibiting the progression of lung cancer.
