Research On The Expression Change Of NLRP3/IL-1? Signaling Pathway In Oral Squamous Cell Carcinoma By LPS Stimulation Of Porphyromonas Gingivalis

Objective:To study the role of lipopolysaccharide(LPS)in Porphyromonas gingivalis(P.g),the main pathogenic bacteria of periodontitis,in the regulation of NLRP3/IL-1? signaling pathway in Oral Squamous Cell Carcinoma(OSCC),and further explore the relevant mechanism of oral microecological imbalance on the occurrence and development of OSCC,and provide new ideas for early prevention and targeted therapy of OSCC.Methods:1.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect mRNA expression of NLRP3,Caspase-1 and IL-1? in adjacent normal and cancer tissues of OSCC patients,normal oral mucosal epithelial cell line(NOK)and OSCC cell lines.2.Western Blot(WB)was used to detect protein expression of NLRP3 and Caspase-1 in adjacent normal and cancer tissues of OSCC patients.3.Enzyme-linked immunosorbent assay(ELISA)was used to detect the protein expression of IL-1? in adjacent normal and cancer tissues of OSCC patients;The protein expression of NLRP3 and Caspase-1 in saliva of OSCC patients and normal people without periodontitis was also detected by it.4.The OSCC cell lines was stimulated by LPS,and mRNA expression of NLRP3,Caspase-1 and IL-1? before and after cell stimulation was detected by qRT-PCR.Results:1.mRNA expression of NLRP3(P=0.053),Caspase-1(P=0.024)and IL-1?(P=0.0362)in OSCC cancer tissues was significantly increased compared with adjacent normal tissues,and the differences were statistically significant,with P values of 0.0353,0.024 and 0.0362,respectively.2.Protein expression in NLRP3,Caspase-1 and IL-1? in OSCC cancer tissues was significantly higher than that in adjacent normal tissues,and the differences were statistically significant,with P values of 0.0354,0.0413 and 0.0148,respectively..3.ELISA showed that the expressions of NLRP3 and Caspase-1 in the saliva of OSCC patients were significantly higher than those of normal people,and the differences were statistically significant,with P values of 0.0106 and 0.0035,respectively..4.Compared with NOK cells,mRNA expression of NLRP3(P<0.0001,P<0.0001),Caspase-1(P<0.0001,P=0.0001)and IL-1?(P=0.003,P=0.0012)in SCC25 and HSC3 cells were significantly increased,and the differences were statistically significant.5.At the same stimulus conditions and concentrations,compared with the control group with equal volume sterile PBS,after 12h of lug/ml LPS stimulation,mRNA expression of NLRP3(P=0.02),Caspase-1(P=0.0176)and IL-1?(P=0.019)in SCC25 cells increased,and the differences were statistically significant.At the same time,the mRNA expression of NLRP3(P=0.0017,P=0.009),Caspase-1(P=0.0109,P=0.0181)and IL-1?(P=0.0005,P=0.016)in HSC3 cells increased after LPS stimulation of lug/ml and 10ug/ml for 48h,and the differences were statistically significant.Conclusions:1.The expression of NLRP3/IL-1? in OSCC cancer tissues is significantly higher than that in adjacent normal tissues,suggesting that it may play an important role in the development of OSCC.2.Stimulation of SCC25 and HSC3 cell lines with LPS of porphyromonas gingival significantly increased the expression of NLRP3/IL-1?.It is suggested that patients with OSCC are accompanied by periodontal disease,leading to oral microecological disorder,which will promote oral cancer.