miR-409-3p调控HMGB1-TLR4通路对胰腺癌细胞增殖迁移及侵袭的影响OA

Objective:To investigate the effects of miR-409-3p on the proliferation,migration,and inva-sion of pancreatic cancer cells,as well as the HMGB1-TLR4 pathway.Methods:qRT-PCR was used to detect the levels of miR-409-3p and HMGB1 mRNA in pancreatic cancer cells.The dual luciferase assay and RNA pull-down assay were used to detect the targeting relationship between miR-409-3p and HMGB1.The PaTu 8988-T cells were divided into the control group,the miR-NC group,the miR-409-3p mimics group,the miR-409-3p mimics+pcDNA3.1 group,and the miR-409-3p mimics+pcDNA3.1-HMGB1 group.Except for the control group,the cells in the other groups were transfected with the corresponding plasmids using Lipo-fectamineTM 3000.qRT-PCR was used to detect mRNA levels of miR-409-3p and HMGB1 in each group of cells.Cell cloning and CCK-8 assay were used to detect cell proliferation.AO/EB staining was used to detect cell apoptosis.Scratch test and Transwell assay were used to detect cell migration and invasion.Western blot was used to detect the expression of proliferation,migration,invasion-related proteins,and proteins related to the HMGB1-TLR4 pathway.The nude mouse tumor transplantation experiment was conducted to detect the effect of miR-409-3p overexpression on the growth of transplanted tumors.Immunohistochemical analysis was conducted to detect the proliferation,migration,and invasion of the transplanted tumors,as well as the expres-sion of proteins related to the HMGB1-TLR4 signaling pathway.Results:In pancreatic cancer cells,miR-409-3p was expressed at a low level,while HMGB1 was expressed at a high level(P<0.05).The RNA pull-down experiment and the dual-luciferase assay revealed a targeting relationship between miR-409-3p and HMGB1.Compared with the miR-NC group,the proportion of apoptotic cells,the level of miR-409-3p in the miR-409-3p mimics group increased,while the number of cell clones,OD450,cell scratch healing rate,cell invasion number,HMGB1 mRNA level,and the expressions of PCNA,c-myc,Snail,MMP-9,HMGB1,and TLR4 decreased(P<0.05).Compared with the miR-409-3p mimics+pcDNA3.1 group,the proportion of apoptotic cells in the miR-409-3p mimics+pcDNA3.1-HMGB1 group decreased,while the number of cell clones,OD450,cell scratch healing rate,cell invasion number,HMGB1 mRNA level,and the expressions of PCNA,c-myc,Snail,MMP-9,HMGB1,and TLR4 increased(P<0.05).The nude mouse tumor transplanta-tion experiment revealed that compared with the miR-NC group,the tumor volume,weight,HMGB1 mRNA level,and the expressions of PCNA,c-myc,Snail,MMP-9,HMGB1,and TLR4 in the miR-409-3p mimics group were all decreased,while the level of miR-409-3p was increased(P<0.05).Conclusion:miR-409-3p may inhibit the proliferation,migration,and invasion of pancreatic cancer cells,and is related to the inhibi-tion of the activation of the HMGB1-TLR4 pathway.