Study on the Mechanism of Jianpi Yangzheng Formula Regulating Tumor-Associated Macrophage Exosomes to Induce Anoikis in Gastric Cancer Cells
10.14148/j.issn.1672-0482.2024.0906
- VernacularTitle:健脾养正方调控肿瘤相关巨噬细胞外泌体诱导胃癌细胞失巢凋亡的机制研究
- Author:
Shanshan ZHENG
1
,
2
;
Jian WU
;
Ruijuan ZHANG
;
Xiang ZHANG
;
Xiaoxia ZHENG
;
Yujia LU
;
Lei HUANG
;
Qingmin SUN
Author Information
1. 南京中医药大学附属医院,江苏 南京 210029
2. 南京中医药大学第一临床医学院,江苏 南京 210023
- Keywords:
gastric cancer;
exosome;
tumor-associated macrophage;
IDH1;
anoikis;
Caspase-3;
tricarboxylic acid cycle metabo-lism
- From:
Journal of Nanjing University of Traditional Chinese Medicine
2024;40(9):906-916
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To investigate the effect of Jianpi Yangzheng(JPYZ)formula regulating tumor-associated macrophage(TAM)exosomes on anoikis in gastric cancer and its mechanism.METHODS TAM model was established by inducing human mononuclear THP-1 cells in vitro;M0,TAM and TAM+JPYZ formula exosomes were extracted by ultracentrifugation and co-incuba-ted with gastric cancer cells.Flow cytometry was used to detect the effect of exosomes in each group on anoikis in gastric cancer cells.A BALB/c transplanted tumor mouse model was constructed,and the expression level of apoptotic proteins in transplanted tumors was detected by Western blot.Label-free mass spectrometry proteomics and bioinformatics were used to analyze the differential proteins in gastric cancer cells before and after intervention;Western blot and qPCR experiments were used to detect the expression level of dif-ferential protein isocitrate dehydrogenase 1(IDH1),and ubiquitination experiments were used to detect the ubiquitination level of IDH1;kits were used to detect the levels of α-ketoglutarate(α-KG),NADPH/NADP+,glutathione(GSH/GSSG)and reactive oxy-gen species(ROS)content.RESULTS The anoikis rate of gastric cancer cells was reduced after TAM exosomes intervention(P<0.05,P<0.01),while it was significantly increased after TAM+JPYZ exosomes intervention(P<0.05,P<0.01).In the in vivo mouse experiment,the ratio of Cleaved Caspase-3/Caspase-3 in the tumor of the TAM exosome group was reduced(P<0.05),while the ra-tio was significantly increased after TAM+JPYZ exosome intervention(P<0.05).Proteomic analysis showed that IDH1 was significant-ly different after intervention,and was related to tricarboxylic acid cycle metabolism.Compared with the M0 group,the IDH1 ubiquiti-nation level of gastric cancer cells in the TAM group with exosome intervention was increased,the levels of α-KG,NADPH/NADP+and GSH/GSSG were significantly increased,and the ROS content was reduced(P<0.05),while TAM+JPYZ exosomes could reverse the above phenomenon.CONCLUSION JPYZ Formula can regulate TAM exosomes to cause ubiquitin degradation of IDH1 in gas-tric cancer cells,reduce the level of tricarboxylic acid cycle metabolism in gastric cancer cells,promote ROS accumulation,induce anoikis in gastric cancer,and thus inhibit the development of gastric cancer.
