Effect Of Porphyromonas Gingivalis On The Proliferation Of Esophageal Squamous Cell Carcinoma

Objective:Esophageal cancer is one of the more common malignant malignancies,with esophageal squamous cell carcinoma(ESCC)predominating in China.The etiology of esophageal cancer is complex,and numerous studies have shown that imbalances in the oral microbiota may be associated with the development of cancers of the digestive tract.In addition,somestudies have demonstrated for the first time the role of oral pathogens in the induction of ESCC tumorigenesis and metastasis.Previous studies by our team involved Porphyromonas gingivalis and its production of toxic components secreted protease-Gingipain may promote ESCC proliferation by activating NF-κB signaling pathway.In this study,we investigated the effect of Porphyromonas gingivalis and its protease secretion on the proliferation of The present study was conducted to investigate the effect of Porphyromonas gingivalis and its protease secretion on the proliferation of esophageal squamous cell carcinoma cells and the mechanism of activation of NF-κB signaling pathway,which provides some reference valuefor the prevention andtreatment ofesophageal squamous cell carcinoma.Methods:At the in vitro cellular level,wild-type Porphyromonas gingivalis,which produces the virulent components secreted protease(Gingipain)and Cathepsin inhibitor 1,and Leupeptin,which replaces the mutation-deficient Porphyromonas gingivalis type KDP136(which does not produce the virulent component Gingipain),were used to infect human esophageal squamous carcinoma Eca109 and KYSE510 cell lines to observe changes in cell biological behavior.Leupeptin,Cathepsin inhibitor 1alonewas used tostimulate Eca109 cells and KYSE30 cells and cell viability was mapped.Results:(1)The CCK8(Cell Counting Kit)assay was performed,and the most appropriate concentrations of Leupeptin and Cathepsin inhibitor 1 were selected for the CCK8 assay to determine cell proliferation and activity;the control group(blank control),model group(wild-type Porphyromonas gingivalis),model+Porphyromonas gingivalis group,model+Cathepsin inhibitor 1group and model+Leupeptin+Cathepsin inhibitor 1 group were set up to treat Eca109 and KYSE510 cells for 24 h and 48 h,model+Leupeptin group,model+Cathepsin inhibitor 1 group and model+Leupeptin+Cathepsin inhibitor 1 group wereused todeterminethe proliferation and activity of Eca109 and KYSE510 cells by CCK8 assay at 24 h,48h,72 h and 96 h,respectively.and KYSE510 cell activities were measured by CCK8 assay at 24 h,48h,72 h and96h.(2)The results of plate cloning experiments showed that the addition of wild Porphyromonas gingivalis alone increased the colony formation of Eca109 cells compared with the control group,Cathepsin inhibitor 1 could inhibit the colony formation of Eca109 cells,Leupeptin had a poor inhibitory effect,and the Model+Cat+Leu group had thebest inhibitory effect on cell colonization.(3)The results of real-time PCR experiments showed that after stimulation of Eca109 cells with wild Porphyromonas gingivalis,KYSE510 cells,the m RNAs of SLC3A2,COPB2 and BIRC2 proteins were more actively expressed in theexperimental group relative tothe blank control group.Conclusion:The present in vitro experiments suggest that Porphyromonas gingivalis maypromote ESCC proliferation through activation ofthe NF-κB signaling.