Abstract: Objective  To investigate the impacts of midazolam (MDZ) on the proliferation,migration,and invasion of esophageal cancer (EC) cells by regulating the monocyte chemotactic protein-1 (CCL2) -CC chemokine receptor 2 (CCR2) signaling pathway.Methods QRT-PCR method was applied to determine the expression of CCL2 and CCR2 mRNA in EC tissue,adjacent cancer tissue,human normal esophageal epithelial cell HEEC,and EC cell Eca-109.MTT assay and colony formation were applied to measure cell proliferation.Scratch test,Transwell test,and TUNEL method were applied to determine cell migration,invasion,and apoptosis,respectively.The expression of CCL2-CCR2 signaling pathway proteins was determined using Western blot method.Results Compared with adjacent cancer tissues and normal human esophageal epithelial cells (HEEC),the mRNA and protein expression levels of CCL2 and CCR2 in cancer tissues and Eca-109 cells were increased (P＜0.05).Compared with the control group,the OD 450 value,colony formation number,scratch healing rate,and invasive cell count of Eca-109 cells in the MDZ-L group,MDZ-M group,and MDZ-H group decreased,while the proportion of TUNEL positive cells increased (P＜0.05).Compared with the MDZ-H group,the OD-450 value,colony formation number,scratch healing rate,and number of invasive cells in the MDZ-H+GW0742 group all increased,while the proportion of TUNEL positive cells decreased (P＜0.05).Compared with the control group,protein and mRNA expressions of CCL2 and CCR2 proteins in Eca-109 cells in the MDZ-L group,MDZ-M group,and MDZ-H group decreased (P＜0.05).Compared with the MDZ-H group,the MDZ-H+GW0742 group showed an increase in the expression of CCL2 and CCR2 proteins in Eca 109 cells (P＜0.05).Conclusion MDZ can inhibit the proliferation,migration,and invasion of EC cells by inhibiting the activation of the CCL2-CCR2 signaling pathway.

Key words: esophageal cancer, midazolam, monocyte chemotactic protein-1, C-C chemokine receptor 2