多梳样蛋白2激活NF-κB信号通路对胶质瘤细胞迁移和侵袭的影响OA

Objective To investigate the effect of polycomb-like protein 2(PCL2)on the migration,invasion,and expression of extracellular matrix(ECM)-related proteins in glioma cells,and to explore its role in glioma invasion.Methods The Cancer Genome Atlas(TCGA)and Chinese Glioma Genome Atlas(CGGA)databases were used to analyze the expression characteristics of PCL2 in gliomas.Ninety glioma tissue specimens were collected from patients admitted to General Hospital of Ningxia Medical University,and the patients'gender,age,and tumor location were recorded.The expression levels of PCL2 in gliomas of different histological grades(Grade Ⅱ,Ⅲ,Ⅳ)were detected by immunohistochemistry.Lentiviral vectors were constructed,and glioma U87-MG cells were divided into control group,PCL2 overexpression group and PCL2 knockdown group.Cell motility,migration,and invasion were assessed by wound healing and Transwell assays;Transcriptome sequencing(RNA-seq)was used to analyze differentially expressed genes(DEGs)and their related functions;Western blotting was employed to determine the expression levels of ECM-related proteins matrix metalloproteinase-7(MMP-7)and MMP-9,as well as nuclear factor-κB(NF-κB)pathway-related transcription factors urokinase-type plasminogen activator(PLAU),interleukin-8(IL-8),and tissue inhibitor of metalloproteinases 2(TIMP-2).Results Analyses of TCGA and CGGA databases revealed that the PCL2 expression level in glioma tissues was significantly higher than that in adjacent non-tumor tissues(P<0.05),and its level in high-grade(Grade Ⅲ and Ⅳ)gliomas was notably higher than that in low-grade(Grade Ⅱ)gliomas(P<0.001).The positive rate of PCL2 expression in glioma patients was 75.6%(68/90),specifically 51.7%(15/29)in Grade Ⅱ,87.5%(14/16)in Grade Ⅲ,and 86.7%(39/45)in Grade Ⅳ.The wound healing and Transwell assays results showed that,compared with control group,PCL2 overexpression group exhibited a increased relative cratch width ratio(P<0.05)and an increased number of migrated cells(P<0.05);PCL2 knockdown group showed an decreased relative scratch width ratio(P<0.05).RNA-seq analysis revealed that 57 DEGs were identified in each of PCL2 overexpression and PCL2 knockdown groups compared with control group,and 119 DEGs were detected between PCL2 overexpression and PCL2 knockdown groups.Gene Ontology(GO)enrichment analysis indicated that the functions of DEGs were enriched in ECM remodeling-related pathways and other processes.Western blotting results showed that,compared with control group,the expression levels of MMP9,MMP7,PLAU,and IL-8 in PCL2 overexpression group were significantly increased(P<0.05),while the expression level of TIMP-2 was decreased(P<0.05);conversely,the expression levels of MMP9,MMP7,PLAU,and IL-8 in PCL2 knockdown group were significantly decreased(P<0.05),while the expression level of TIMP-2 was increased(P<0.05).Conclusion PCL2 promotes the migration and invasion abilities of glioma cells,which may be associated with the activation of the NF-κB signaling pathway.