Abstract：Objective: To investigate the impact and mechanism of erianin on the malignant progression of esophageal cancer (EC) cells. Methods: The cancer tissues and adjacent tissues 3cm away from the cancer tissues of EC patients (40cases) who underwent radical surgery in our hospital from March 2021 to December 2022 were collected. Real-time fluorescence quantitative PCR (qRT-PCR) and western blot were applied to detect the mRNA and protein expression levels of CCL2 and CCR2 in EC tissues/cells, respectively; ECA109 cells were treated with different concentrations of erianin (0, 10, 20, 40, 60, 80, 160 nmoL/L) for 48 h, and cell proliferation was detected by MTT assay to screen the optimal intervention concentration. ECA109 cells were divided into control group (normal culture), low, medium and high dose erianin groups (20,40,60nmol/L), pcDNA group (transfected with pcDNA3.1), and pcDNA-CCL2 group (transfected with pcDNA3.1-CCL2), MTT assay, plate cloning assay, transwell chamber and flow cytometry were applied to detect cell proliferation, migration, invasion and apoptosis, respectively; after co-culturing ECA109 cells and CD8+T cells, they were grouped into T cell group, co-culture group, erianin group, pcDNA group, and pcDNA-CCL2 group. MTT and flow cytometry were applied to detect CD8+T cell proliferation and apoptosis, enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of interferon-γ (IFN-γ), interleukin-2 (IL-2), and tumor necrosis factor-α (TNF-α) in the supernatant of CD8+T cells; Western blot was applied to detect the expression of proliferating cell nuclear antigen (PCNA), Bcl-2 associated protein (Bax), programmed death ligand 1 (PD-L1), and CCL2-CCR2 signaling pathway protein in cells. Results: The expression levels of CCL2, CCR2 proteins, and mRNAs increased in EC tissues and EC cells (P<0.05); compared with the control group, the OD490 value, colony formation number, migration and invasion cell numbers, CCL2, CCR2 mRNA, PCNA, PD-L1, CCL2, and CCR2 protein expression levels in ECA109 cells in the low, medium, and high dose erianin groups were obviously reduced, the apoptosis rate and Bax protein expression level were obviously increased (P<0.05); compared with the pcDNA group, the changes in the above indicators of ECA109 cells in the pcDNA-CCL2 group were obviously reversed (P<0.05); compared with the T cell group, the OD490 value, IFN-γ, IL-2, and TNF-α levels of CD8+T cells in the co-culture group decreased, while the cell apoptosis rate increased (P<0.05); compared with the co-culture group, the OD490 value, IFN-γ, IL-2, and TNF-α levels of CD8+T cells in the erianin group increased, while the cell apoptosis rate decreased (P<0.05); compared with the pcDNA group, the changes in the above indicators of CD8+T cells in the pcDNA-CCL2 group were obviously reversed (P<0.05). Conclusion: Erianin may inhibit the malignant progression of EC cells by inhibiting the CCL2-CCR2 signaling pathway.
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2023, Vol. 29 
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