Resveratrol Activates MAPK Pathway to Regulate Estrogen Metabolism in Type I Endometrial Cancer
Authors: Wang Qing , Zhou Jia-Yun , Zhang Yong-Li , Yin Ze-Yuan , Liu Li , Lu Hui , Zhou Xue-Yan , Zhang Bei 
Published: February 2023

Abstract
Purpose
Endometrial cancer(EC) is an estrogen-dependent tumor, the occurrence of which is closely related to an imbalance of estrogen homeostasis. Our previous studies explored the effects of resveratrol on estrogen metabolism.However, systematic research on the exact mechanism of action of resveratrol is still lacking.Based on network pharmacology, molecular docking and animal experiments, the effects and molecular mechanisms of resveratrol on endometrial cancer were investigated.
Methods
The target of resveratrol was obtained from HERB and ECTM databases, and the target of endometrial cancer was obtained by using Genecards database.Use of Veeny map to obtain the intersection target of resveratrol in the treatment of EC, and the protein interaction network of the intersection target was constructed by importing the STRING database,then construct the drug-disease-target interaction network based on Cytoscape 3.9.1 software.GO and KEGG pathway enrichment analysis were performed for intersection targets using OmicShare cloud platform. Resveratrol and core target were analyzed by molecular docking. EC mice model induced by MNNG were randomly divided into Control group, Res group, MNNG group, and MNNG + Res group. MNNG + Res + MAPK/ERKi group.The protein expression levels of ERK and p-ERK in mouse uterus were detected by Western blot.The contents of E1, E2, E3, 16-epiE3, 17-epiE3, 2-MeOE1, 4-MeOE1, 2-MeOE2, 4-MeOE2, 3-MeOE1, 2-OHE1, 4-OHE1, 2-OHE2, 4-OHE2, 16α-OHE1 in serum and endometrial tissue of mice were detected by LC-MS/MS.
Results
A total of 174 intersection targets of resveratrol anti EC were obtained. Signaling pathways analyzed by KEGG enrichment included AGE-RAGE signaling pathway in diabetic complications, PI3K-Akt signaling pathway and MAPK signaling pathway.The top 10 core targets were MAPK3, JUN, TP53, CASP3, TNF, IL1B, AKT1, FOS, VEGFA and INS.Molecular docking showed that except TNF, other targets had good affinity with resveratrol, and the binding activity with MAPK3 was stable.Western blot results showed that resveratrol increased the phosphorylation level of ERK, and MAPK/ERKi decreased the activation of ERK.Using LC-MS/MS analysis,the contents of 2-MeOE1,2-MeOE2 and 4-MeOE1 in serum and uterine tissue were showed significantly decreasing trend in MNNG group, while the content of 4-OHE2 was increased (P༜0.05).The concentration levels of 4-MeOE1 in serum and 2-MeOE1 and 2-MeOE2 in endometrial tissue of mice were significantly increased after resveratrol treatment,the content of 4-OHE2 in serum and uterus of mice was significantly decreased (P༜0.05). Meanwhile, MAPK/ERKi intervention group showed that the effect of resveratrol on the reversal of estrogen homeostasis imbalance was obviously weakened.
Conclusion
Resveratrol has the characteristics of multiple targets and multiple approaches in the treatment of endometrial cancer. In this study, it was found that resveratrol regulates estrogen metabolism by activating MAPK pathway, which provides a new perspective for subsequent research on the treatment of endometrial cancer.