MicroRNA-155-5p targets caudal-type homeobox 1 to inhibit the migration,invasion and epithelial-mesenchymal transition of human pancreatic cancer cells
Objective To investigate the role of caudal-type homeobox 1(CDX1)in the migra-tion,invasion and epithelial-mesenchymal transition(EMT)of pancreatic cancer(PC)cells,and the mo-lecular mechanism of microRNA-155-5p(miR-155-5p)/CDX1 axis in regulating the migration,invasion and EMT of PC cells.Methods Cancer tissues from 30 patients with PC who underwent PC resection in Linyi People's Hospital from January 2020 to January 2023 were selected as the research objects.Immuno-histochemical staining,Western blotting and real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)were used to detect the expression level of CDX1 in PC tissues.Cell counting kit-8(CCK-8)assay,wound healing assay and Matrigel invasion assay were used to detect cell migration and invasion a-bility.Western blotting and RT-qPCR were used to detect the expression levels of EMT-related genes E-cadherin,N-cadherin and Vimentin.Luciferase reporter gene assay was used to detect the binding of miR-155-5p to CDX1 promoter region.The differences between groups were analyzed by t-test or analysis of variance(ANOVA).Results The expression levels of CDX1 protein and RNA in PC tissues(7.15± 1.72,9.17±2.83)was significantly higher than that in adjacent tissues(1.00±0.28,1.00±0.38)(t=11.290,13.912,P＜0.05).Overexpression of CDX1 promoted the migration and invasion of PC cells.Scratch test showed that the number of migrating cells in overexpression(Ad-CDX1)group was sig-nificantly higher than that in control(Ad-GFP)group(81.92±5.08 vs.53.10±4.19,t=10.031,P＜0.01).Overexpression of miR-155-5p inhibited the migration and invasion of PC cells,and the number of migrating cells in miR-155-5p mimics group was significantly less than that in negative control(NC)mim-ics group(38.80±5.76 vs.53.48±6.19,t=7.861,P＜0.01).The results of RT-qPCR showed that the expression level of EMT-related gene E-cadherin in miR-155-5p inhibitor+small interfering RNA(si)-CDX1 group was higher than that in miR-155-5p inhibitor+si-NC group(1.73±0.36 vs.1.00± 0.12,t=6.001,P＜0.05),and the expression levels of N-cadherin and Vimentin were lower than those of miR-155-5p inhibitor+si-NC(N-cadherin:1.16±0.21 vs.3.18±0.67,t=4.337,P＜0.05;Vim-entin:1.23±0.41 vs.2.76±0.54,t=6.230,P＜0.05).The results of Western blotting and luciferase reporter gene assay indicated that miR-155-5p directly targeted CDX1 gene to exert the above effects.Conclusion The expression of CDX1 is significantly increased in PC cells,and miR-155-5p inhibits the migration,invasion and EMT of human PC cells by directly targeting CDX1.
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