Effects of Yijing Buqi Yangxue prescription on the proliferation and related signal transduction of esophageal cancer cells by activating T cells.

e16122 Background: Esophageal cancer is a malignant disease that seriously threatens human health. This study aims to screen out the effective Chinese herbs with strong inhibitory effects on the proliferation of esophageal cancer(EC) cells EC-1, Eca109, EC9706 though regulating immune cell, and further study the signal transduction mechanism of composed Yijing Buqi Yangxue Prescription(YBYP). Methods: The T cell activation model was constructed by Jurkat cell line, and the Chinese herbs that can inhibit the proliferation of EC cells by activating T cells was screened from 45 kinds of Chinese herbs with the effect of trengthening healthy qi. Orthogonal design test was used to determine the optimal dosage ratio of compound Chinese medicine prescription, and composing YBYP. Then transcriptome sequencing, bioinformatics, RT-qPCR and Western blot methods were applied to study the signal transduction mechanism of T cells activated by Yijing Buqi Yangxue prescription. Results: T cell activation model was successfully established while Jurkat cells were stimulated by 2μg/mL PHA-M and 200 nm PMA for 24h, and the expressions of CD25, IL-2 and IFN-γ increased significantly. When the activated Juakat cells were co-cultured with EC cells at 2.5:1 for 48h, the inhibitory effect of Juakat cells on EC cells increased significantly, reaching 70%. Semen Cuscutae (Tusizi), Angelica Sinensis (Danggui), Glycyrrhiza Uralensis Fisch (Gancao) were screened out and composed YBYP according to their efficacy and orthogonal design. YBYP could obviously inhibit the proliferation of EC9706, EC-1, ECa109 cells with dose-effect manner. IC50 values of Tusizi, Danggui, Gancao in YBYP for EC9706 cell were 49.072μg/ml, 88.768μg/ml, 148.887μg/ml, respectively; for EC-1 cell were 19.197μg/ml, 13.762μg/ml, 8.109μg/ml respectively; for ECa109 cell were 10.548μg/ml, 16.706μg/ml, 17.342μg/ml respectively. YBYP colud significantly up-regulate the protein expressions of NOTCH1, RBP-Jκ, HSPB1, MAML1, RUNX3 and down-regulate the expression of GATA3 to promote the proliferation and activation of Jurkat cells, and regulate the protein and mRNA expression of cMyc/RUNX3/AKT1, EGFR/ERK/cFos and JAK2/STAT3/Bcl-2 signaling pathways of EC cells EC9706, EC-1 and Eca109. Conclusions: YBYP could enhance the inhibitory effect on the proliferation of EC cells EC9706, EC-1 and Eca109 by activating T cells through HSPB1/NOTCH1 (ICN) /MAML1 related signaling pathway, and regulate cMyc/RUNX3/AKT1 EGFR/ERK/cFos and JAK2/STAT3/Bcl-2 signaling pathways.