AIM2对宫颈癌细胞PANoptosis的影响及其机制OA

Objective To investigate the effect of AIM2 on PANoptosis in cervical cancer HeLa cell and its underlying mechanism.Methods ①The expression level of AIM2 gene in cervical cancer was analyzed using UCSC database.Western blot and RT-qPCR were performed to detect the expressions of AIM2 and PANoptosis-related molecules[pyroptosis(Caspase-1),apoptosis(Caspase-3,cleaved-Caspase-3),necroptosis(MLKL,p-MLKL)]in cervical cancer cells HeLa,C33a and normal cervical epithelial HCer cells.②Lentiviral transfection was used to establish a stable AIM2-knockdown HeLa cell model(sh-AIM2),and blank group and negative control group(sh-NC)were set up in the experiment.Western blot was used to detect the expressions of AIM2 and PANoptosis-related pro-teins.Pyroptosis,apoptosis and necroptosis were detected by EthD-Ⅲ,TUNEL and PI staining,respectively.③The expression level of PLA2G7 gene in cervical cancer cell and its correlation with AIM2 expression were analyzed based on UCSC and GEPIA databases,respectively.Western blot was used to detect the expression of PLA2G7 in cervical cancer cells and its change after AIM2 knockdown.④HeLa cells were divided into control group,Melittin(PLA2G7 agonist)group,sh-AIM2 group,and sh-AIM2+Melittin group.Western blot was used to detect the expressions of AIM2,PLA2G7 and PANoptosis-related molecules.Results ①Based on UCSC database,AIM2 expression was significantly higher in cervical cancer tissues than in normal cervical epithelial tissues(P<0.05).Compared with HCer and C33a cells,HeLa cells exhibited increased protein and mRNA levels of AIM2,elevated mRNA levels of Caspase-1,Caspase-3 and MLKL,and increased protein expressions of Caspase-1,cleaved-Caspase-3 and p-MLKL(all P<0.05).②Compared with blank group,AIM2 protein expression,the expressions of Caspase-1,cleaved-Caspase-3 and p-MLKL,and the levels of pyroptosis,apoptosis and necroptosis were significantly reduced in sh-AIM2 group(P<0.05).③UCSC database analysis results showed that PLA2G7 expression was significantly higher in cervical cancer tissues than in normal cervical epithelial tissues(P<0.05).GEPIA database analysis results showed that there was a positive correlation between AIM2 expression and PLA2G7 expression in cervical cancer patients(r=0.27,P<0.05).Western blot results indicated that PLA2G7 expression was significantly higher in HeLa cells than in C33a cells,and that PLA2G7 expression was significantly decreased after AIM2 down-regulation(P<0.05).④Compared with control group,the protein expressions of PLA2G7,Caspase-1,cleaved-Caspase-3 and p-MLKL in HeLa cells were up-regulated in Melittin group(P<0.05),while these molecules were significantly down-regulated in sh-AIM2 group(P<0.05).Compared with sh-AIM2 group,the low expression of these molecules were reversed in sh-AIM2+Melittin group(P<0.05).Conclusion AIM2 and PLA2G7 are highly expressed and positively correlated in cervical cancer.Down-regulation of AIM2 can reduce PLA2G7 expression,and they synergistically promote PANoptosis in HeLa cells,providing a potential direction for cervical cancer treatment.