Effect and mechanism of Yangyin Huowei Mixture(YYHWM)down-regulating Aquaporin 3(AQP3)on MC cell phenotype based on single-cell sequencing data
Objective To observe the effect of Yangyin Huowei Mixture(YYHWM)on MC cell(MNNG induces malignant transformation of GES-1 in human gastric mucosal epithelial cells)proliferation,migra-tion,invasion and apoptosis,and to explore the mechanism of its downregulating AQP3 to inhibit the acti-vation of IL-10/JAK1/STAT3 signaling pathway to block or reverse CAG lesions.Methods Selected CAG single cell transcriptome sequencing data from GEO database and drawn the expression matrix of the data.UMAP visualization was processed using Seurat 4.3.0 package of R language.MC cells were divided into YYHWM group,Lentiviral transfection group with low expression of AQP3(AQP3 low-expression group),YYHWM and Lentiviral transfection group with high expression of AQP3(YYHWM+AQP3 high-expression group).MC cells and normal human gastric mucosa epithelial cells GES-1 were used as model control group(MC group)and blank control group(control group).Cell phenotype changes were detected by plate cloning,scratch assay,transwell and flow cytometry.IL-10 expression levels were deter-mined by ELISA and the expression levels of JAK1 and STAT3 were determined by western blot.Res-ults Analysis of single-cell sequencing datasets revealed the expression level of AQP3 elevated in CAG samples and epithelial cell populations.AQP3 may affect the pathological process of precancerous GC through IL-10 anti-inflammatory signaling pathway.Compared with MC group,the AQP3 low-expression group and YYHWM group showed reduced cell invasion,migration and proliferation,increased apoptosis rate,decreased IL-10 levels in the cell culture supernatant,and decreased JAK1 and STAT3 protein ex-pression levels in the cells(P<0.05 or P<0.01),YYHWM+AQP3 high-expression group showed no significant differences(P>0.05).Conclusion YYHWM can inhibit the activation of IL-10/JAK1/STAT3 signaling pathway by down-regulating the expression of AQP3,thus weakening the invasion,mi-gration and proliferation of MC cells,promoting cell apoptosis and thus blocking or reversing the patholog-ical process of CAG.
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