Mechanism of Weichang' an on Autophagy of MKN45 Cells
10.13422/j.cnki.syfjx.20192323
- VernacularTitle:胃肠安诱导胃癌MKN45细胞自噬的机制
- Author:
Zhao-yan LI
1
;
Wei-xia CHEN
2
;
Meng-meng QIN
1
;
Jia LI
1
;
Yan XU
1
;
Ai-guang ZHAO
1
Author Information
1. Longhua Hospital, Shanghai University of Traditional Chinese Medicine (TCM), Shanghai 200032, China
2. Henan Province Hospital of TCM, Zhengzhou 450002, China
- Publication Type:Research Article
- Keywords:
invigorating spleen;
Weichang'an;
gastric cancer;
autophagy;
MKN45 cells
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2020;26(1):71-77
- CountryChina
- Language:Chinese
-
Abstract:
Objective::To explore the effect of Weichang' an (WCA) on the autophagy of human gastric cancer MKN45 cells and its possible anti-cancer mechanism. Method::MKN45 cells were cultured in vitro and incubated with different concentrations (250, 500, 1 000, 2 000 mg·L-1) of WCA for 24, 48, 72 h. Cell counting kit-8 (CCK-8) assay was used to detect the cell proliferation. AO/EB Dyeing (AO) staining and monodansylcadaverin (MDC) staining were used to observe the changes of the effect of WCA on autophagosome and autophagic vesicles in gastric cancer MKN45 cells at 48 h. Real-time polymerase chain reaction (Real-time PCR) and Western blot were used to detect microtubule-associated protein 1 light chain 3 (LC3), Beclin1, sequestosome 1 (p62), human autophagy-related gene 5 (ATG5), human autophagy-related gene 7 (ATG7) mRNA and protein expression levels. Result::WCA showed a dose-and-time-dependent growth inhibition at the concentration above 1 000 mg·L-1. Compared with the blank group, WCA (500, 1 000, 2 000 mg·L-1) significantly inhibited the proliferation of MKN45 cells (P<0.05, P<0.01). AO staining and MDC staining showed that autophagosomes and autophagic sacs increased with the rise of WCA concentration compared with the blank group. Real-time PCR and Western blot showed that the expressions of autophagy-related proteins LC3-Ⅱ, Beclin1, ATG5, ATG7, mRNA and protein increased gradually after WCA (1 000 mg·L-1) intervention, while p62 expression decreased (P<0.05, P<0.01). Conclusion::WCA induces the autophagy of human gastric cancer MKN45 cells in a time-and dose-dependent manner in vitro, which may be related to the up-regulation of LC3-Ⅱ, Beclin1, ATG5 and ATG7 as well as the down-regulation of p62 and LC3-Ⅰ.
