Molecular mechanisms of the Xiao-chai-hu-tang on chronic stress-induced colorectal cancer growth based on an integrated network pharmacology and RNA sequencing approach with experimental validation

Abstract Background Chronic stress is a risk factor for the development of colorectal cancer (CRC). Xiao-Chai-Hu-Tang (XCHT) is a traditional Chinese medicine prescription and has been widely used to treat chronic stress-related diseases and cancer. However, its role in chronic stress-induced CRC remains unclear. Methods Our study aimed to investigate the roles of XCHT in CRC development under chronic stress. A xenografted CRC mouse model exposure to chronic restraint stress (CRS) was used to determine the effects of XCHT on CRC growth in vitro and in vivo. XCHT (10.27, 20.54 g/kg) were given by oral gavage once a day. RNA-sequencing was combined with network pharmacology to investigate potential target and pathway in this study. ELISA, RT-qPCR and immunofluorescence were performed to detect the expression of inflammation related genes. Glycolysis related genes and phenotype were evaluated by WB, RT-qPCR and seahorse. Results XCHT significantly inhibited CRC growth under chronic stress in a dose-dependent manner. Mechanistically, XCHT suppressed the expression levels of glycolysis associated enzymes and inflammatory factors caused by chronic stress. Moreover, XCHT significantly mitigated the activity of the JAK2/STAT3 signaling which was activated by chronic stress induced IL-6. Conclusions XCHT could reverse the tumor energy metabolism reprogramming and improve the inflammatory microenvironment in CRC under chronic stress through the IL-6/JAK2/STAT3 pathway. Therefore, XCHT might represent a promising therapeutic strategy for suppressing psychologically associated CRC progression.