Virtual Screening and Reality Verification:Elemene Injectable Emulsion acts on the key targets and pathways of Colorectal Adenoma Cancerization

Abstract

Purpose Most colorectal cancer(CRC) is developed from intestinal adenomatous polyps. Therefore, it is urgent to find new therapeutic drugs to intervene intestinal adenoma development in CRC.ELEMENE INJECTABLE EMULSION(EIE) has been reported to exert antitumor activity in various digestive tumor diseases. However, the mechanism of EIE in preventing colorectal adenoma (precancerous lesions) from developing into CRC has not been systematically explored.Methods Using network pharmacology correlation analysis and molecular docking, the central target of EIE in preventing colorectal adenoma(CRA) from transforming into cancer through innate immunity was excavated and verified. The differentially enriched pathways of human CRA, CRC, and corresponding adjacent tissue samples were analyzed by reverse-phase protein array (RPPA) to verify the relevant mechanism. Colon cancer cells were intervened to observe the proliferation, apoptosis, and cell cycle in different concentrations of EIE. The predicted related targets were verified by RT-PCR(real-time PCR), and the pathways were confirmed by Western blot.Results The analysis results show that Retinoid X Receptor alpha (RXRa) was the key target gene, and the main pathway was PI3K/AKt. Molecular docking results show that β- Elemene,γ-Elemene, and δ- Elemene have a strong affinity for RXRa.RPPA technology was used to analyze the functional enrichment of the differentially expressed genes of the Adenoma Cancer sequence, Adenoma Paracancerous sequence, and Cancer-Paracancerous sequence. The enrichment results of the three groups of sequence differential genes showed that the PI3K/Akt signaling pathway was the most significant. In addition, based on HCT116 and THC8307 in vitro experiments, PI3K,p-PI3K, Akt,p-Akt, and RXRa proteins and the relative expression of RXRa mRNA in the EIE intervention group were studied, and the predicted results were verified.Conclusion This is also the first evidence that our data provide that elemene aims to target the PI3K-Akt signaling pathway and RXRa, a target gene to play its role in affecting the development of CRA in cancer through innate immunity.