Abstract 1030: Effects of clearing heat and removing phlegm prescription on inhibiting proliferation and secreted protein expression of human esophageal carcinoma cells

Abstract Background: Esophageal carcinoma(EC) is the sixth most common cause of cancer death worldwide. It is still a key problem to find early diagnostic targets and effective treatment medicine in the study of EC. This study aims to screen out serum differentially expressed proteins from EC patients, and explore the inhibitory effects of Clearing Heat and Removing Phlegm prescription(CHRF) on the proliferation of EC cells and serum secretory proteins expression. Methods: Serum samples of 21 EC patients and 7 healthy normal people were collected. Raybiotech AAH BLG-1 protein chip and data analysis software were used to profile serum differentially expressed proteins of EC. CHRF was developed in our previous research for the treatment of EC. Effects of CHRF on the proliferation, cell cycle and screened serum secretory protein expression of four EC cell lines EC-1, Eca109, EC9706, TE-1 were investigated by MTT assay, flow cytometry assay and Western blot analysis. Results: 37 differentially expressed proteins were screened out from EC patients compared with the normal control group, among which 22 were significantly up-regulated, 17 were significantly down-regulated. These differential expression proteins mainly involved in cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, Jak-STAT signaling pathway, chemokine signaling pathway, which have the function of promoting tumor development, tumor growth, angiogenesis, growth and development of tissue. 22 up-regulated serum secretory proteins were selected to detect in four EC cells and observe the inhibitory effects of CHRF. CHRF could obviously inhibit the proliferation of EC-1, EC9706, Eca109 and TE-1 cells in dose-dependent and time-dependent manner. IC50 values of CHRF for EC9706, Eca109, TE-1 and EC-1 were 33.31μg/ml, 20.70μg/ml, 21.93μg/ml, 33.47μg/ml respectively. CHRF induced G2/M phase arrest in all four EC cells. MMP1, MMP14, OPG, FBLN5, PECAM-1 proteins were expressed in all four EC cells and down-regulated by CHRF. Conclusion: Serum secretory proteins MMP1, MMP14, OPG, FBLN5, PECAM-1 were significantly up-regulated in esophageal cancer cells, which could be considered as the potential markers for EC diagnosis; the inhibitory effects of CHRF on the proliferation of Eca109, EC9706, TE-1 and EC-1 cells have relationship with down-regulating MMP1, MMP14, OPG, FBLN5 and PECAM-1 protein expression, which not only provides new basis for the in-depth study of EC pathogenesis, but also provides foundation for the further study of new traditional Chinese medicine prescriptions for EC treatment. Citation Format: Fuchun Si, Gao Si, Xuejie Song. Effects of clearing heat and removing phlegm prescription on inhibiting proliferation and secreted protein expression of human esophageal carcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1030.