Experimental study on the promotion of lung adenocarcinoma metastasis by IRF6 activation of MMP-12 transcription
Objective To investigate the effect of interferon regulatory factor 6(IRF6)on lung adenocarcinoma(LUAD)cell metastasis and its possible mechanism.Methods The expression of IRF6 in LUAD tissues and normal lung tissues was obtained from GEPIA and Ualcan online databases.Real-time quantitative fluorescent PCR(qPCR)was used to detect the expression of IRF6 and matrix metallopeptidase 12(MMP-12)in LUAD cells and normal bronchial epithelial cells.LUAD cells were transfected with sh-NC and sh-IRF6 respectively,and the migration ability of the two groups of cells was detected by Transwell assay.MMP-12 promoter enrichment was detected by chromatin immunoprecipitation(ChIP)assay.The targeting relationship between IRF6 and MMP-12 was verified by luciferase reporter gene assay.Results The result of online database analysis showed that the expression of IRF6 in LUAD tissues was significantly higher than that in normal tissues.Compared with normal cells(BEAS-2B),the expression of IRF6 in A549 and Calu3 cells was significantly increased(5.82±0.92,7.11±1.31,P<0.01).Compared with sh-NC group,the migration numbers of A549 and Calu3 cells in sh-IRF6 transfection group were decreased(52±9.23 and 63±11.38 vs.21±4.24 and 33±6.11),and the difference was statistically significant(P<0.01).The expression of MMP-12 in A549 and Calu3 cells decreased to 0.31±0.06 and 0.45±0.11 in sh-IRF6 transfection group,and the difference was statistically significant(P<0.01).The ChIP test result showed that compared with the negative Anti-IgG group,the MMP-12 promoter enrichment of A549 and Calu3 cells in Anti-IRF6 group was increased by(31.24±5.78)fold and(26.27±4.11)fold,and the difference was statistically significant(P<0.01).After interference with IRF6,the transcriptional activity of MMP-12 in A549 and Calu3 cells was significantly reduced to 0.28±0.04 and 0.38±0.06(P<0.01).The overexpression of MMP-12 in IRF6-interfering A549 and Calu3 cells increased the cell migration numbers from 25±4.21 and 36±7.28 to 48±8.89 and 60±11.17(P<0.01).Conclusion IRF6 is highly expressed in LUAD cells,and knocking down IRF6 expression can significantly inhibit LUAD cell metastasis.
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